Prevalence and risk factors of Hepatitis C virus infection in Brazil, 2005 through 2009: a cross-sectional study

  • Leila MMB Pereira1, 2,

    Affiliated with

    • Celina MT Martelli3, 4,

      Affiliated with

      • Regina C Moreira5,

        Affiliated with

        • Edgar Merchan-Hamman6,

          Affiliated with

          • Airton T Stein7,

            Affiliated with

            • Regina Maria A Cardoso8,

              Affiliated with

              • Gerusa M Figueiredo8,

                Affiliated with

                • Ulisses R Montarroyos1, 4,

                  Affiliated with

                  • Cynthia Braga9,

                    Affiliated with

                    • Marília D Turchi3,

                      Affiliated with

                      • Gabriela Coral7,

                        Affiliated with

                        • Deborah Crespo10,

                          Affiliated with

                          • Maria Luiza C Lima9, 11,

                            Affiliated with

                            • Luis Claudio A Alencar1,

                              Affiliated with

                              • Marcelo Costa12,

                                Affiliated with

                                • Alex A dos Santos13 and

                                  Affiliated with

                                  • Ricardo AA Ximenes1, 4Email author

                                    Affiliated with

                                    BMC Infectious Diseases201313:60

                                    DOI: 10.1186/1471-2334-13-60

                                    Received: 6 August 2012

                                    Accepted: 11 January 2013

                                    Published: 1 February 2013

                                    Abstract

                                    Background

                                    Hepatitis C chronic liver disease is a major cause of liver transplant in developed countries. This article reports the first nationwide population-based survey conducted to estimate the seroprevalence of HCV antibodies and associated risk factors in the urban population of Brazil.

                                    Methods

                                    The cross sectional study was conducted in all Brazilian macro-regions from 2005 to 2009, as a stratified multistage cluster sample of 19,503 inhabitants aged between 10 and 69 years, representing individuals living in all 26 State capitals and the Federal District. Hepatitis C antibodies were detected by a third-generation enzyme immunoassay. Seropositive individuals were retested by Polymerase Chain Reaction and genotyped. Adjusted prevalence was estimated by macro-regions. Potential risk factors associated with HCV infection were assessed by calculating the crude and adjusted odds ratios, 95% confidence intervals (95% CI) and p values. Population attributable risk was estimated for multiple factors using a case–control approach.

                                    Results

                                    The overall weighted prevalence of hepatitis C antibodies was 1.38% (95% CI: 1.12%–1.64%). Prevalence of infection increased in older groups but was similar for both sexes. The multivariate model showed the following to be predictors of HCV infection: age, injected drug use (OR = 6.65), sniffed drug use (OR = 2.59), hospitalization (OR = 1.90), groups socially deprived by the lack of sewage disposal (OR = 2.53), and injection with glass syringe (OR = 1.52, with a borderline p value). The genotypes 1 (subtypes 1a, 1b), 2b and 3a were identified. The estimated population attributable risk for the ensemble of risk factors was 40%. Approximately 1.3 million individuals would be expected to be anti-HCV-positive in the country.

                                    Conclusions

                                    The large estimated absolute numbers of infected individuals reveals the burden of the disease in the near future, giving rise to costs for the health care system and society at large. The known risk factors explain less than 50% of the infected cases, limiting the prevention strategies. Our findings regarding risk behaviors associated with HCV infection showed that there is still room for improving strategies for reducing transmission among drug users and nosocomial infection, as well as a need for specific prevention and control strategies targeting individuals living in poverty.

                                    Keywords

                                    Hepatitis C virus Prevalence Genotype Cross sectional study Risk factors

                                    Background

                                    Around the world, infection by the Hepatitis C virus (HCV) causes acute and chronic liver disease and may lead to cirrhosis, liver failure and/or hepatocellular carcinoma. HCV chronic liver disease is one of the main causes of liver transplantation in developed countries [1]. The global prevalence of HCV infection is estimated to be 2.2%, with 170 million individuals infected (2004) [2].

                                    The sequential population-based studies conducted in the USA are among the few nationwide surveys to estimate the prevalence of HCV [36]. In the USA, a 1.6% nationwide prevalence of anti-HCV was found by the National Health and Nutrition Examination Survey Continuous (NHANES Continuous) between 1999 and 2002 [5]. These surveys showed a decline in the prevalence of hepatitis C from NHANES III (1988–1994) to NHANES Continuous (2003–2006), but this figure remained stable between the NHANES 1999–2002 and NHANES 2003–2006 [6]. The incidence of acute hepatitis C tended to decline over the twenty-five years of surveillance (1982–2006) in selected counties in the USA, with intravenous drug use (IDU) continuing to be the predominant risk factor [7]. In France, a population-based survey showed an overall prevalence of anti-HCV of 0.84% in 2004. IDU, nasal drug use, blood transfusion prior to 1992, having a tattoo, being older than 29 years, social deprivation, and being born on a continent with endemic levels higher than 2.5% were the risk factors associated with HCV infection [8].

                                    In Latin America, estimates of HCV prevalence range from 1.0 to 2.3% using projections of HCV for modeling [9]. Data from the World Health Organization (WHO) placed Brazil among the countries with a prevalence of hepatitis C infection ranging from 2.5% to 10% [2]. Compulsory notification of acute and/or chronic hepatitis C was implemented by the Brazilian surveillance system in 1996. The detection rates for anti-HCV remained stable at approximately 10 cases per 100,000 individuals from 2004 to 2009, with the bulk of cases being reported in the southeast region [10]. In Brazil, the few population-based studies [1113] and the vast majority of studies reporting risk factors associated with HCV infection were obtained from selected populations in restricted areas [1418]. In a large sample of chronic HCV patients from different laboratories located in all Brazilian macro-regions, genotype 1 was found to predominate, followed by genotypes 3, 2, 4 and 5 [19].

                                    A national seroepidemiological survey of Hepatitis A, B and C infection was launched by the Brazilian Ministry of Health in 2005 and completed in 2009. The epidemiology of hepatitis A and hepatitis B for the northeast and central-west regions and the Federal District has already been published [20, 21]. The aim of the present study was to estimate the seroprevalence of HCV antibodies in the five macro-regions of Brazil and the Federal District. The association between potential risk factors and HCV seropositivity was assessed for the country as a whole and the population attributable risk for multiple factors was estimated using the case–control approach. This is the first population-based HCV antibody survey conducted in a representative sample of the Brazilian macro-regions and the Federal District.

                                    Methods

                                    Study design and sites

                                    The national seroepidemiological survey of hepatitis A, B and C infection was designed as a representative sample of individuals aged 10 to 19 and 20 to 69 years in the 5 Brazilian macro-regions. Participants were residents of all 26 State capitals and the Federal District. The fieldwork was conducted from 2005 to 2009. The methodology has been described in a prior publication [22].

                                    In brief, a stratified multistage cluster sampling method based on The Brazilian Institute of Geography and Statistics (IBGE) census data for the year 2000 was used to select the study population. In the first step, within each macro-region, the sample was stratified by state capital using a constant sampling fraction. In the second step the primary sampling units – census tracts – were drawn with probability proportional to population size and assuring the inclusion of all socioeconomic groups. The primary sampling units were subsampled successively in terms of blocks (with probability proportional to population size), households and eligible individuals. The final analysis weights incorporated the selection probabilities and design effect due to the cluster sampling to obtain representative estimates of the target population.

                                    Data collection and variables

                                    All sites followed the same standard protocol. Data were collected by interview using a structured questionnaire during household visits. Blood samples were collected after the interviews. Both procedures were performed by trained health personnel.

                                    The individual variables collected were social-demographic characteristics, blood route-related factors, such as dental treatment, surgery, history of blood transfusion, hospitalization, endoscopy, use of glass syringe not related with drug abuse, tattoos, or body piercing. Questions related to alcohol, drug consumption, lifelong sexual behavior and health-related jobs were restricted to persons aged 13 years or older. At the household level, data on income and education of the head of family and the characteristics of the household including home ownership, water supply, sewage and solid waste disposal were recorded [22].

                                    Serum tests and definitions

                                    Blood samples were collected, transported and stored according to standard procedures. HCV antibodies were detected by a third-generation (Axsym, Abbott Laboratories, Abbott Park, Illinois) enzyme immunoassay in central public health laboratories. The results were considered positive when the optical density (OD) was ≥ 1.2 (20% higher than the cutoff). Samples with OD between 1.0 and 1.19 were considered indeterminate and were retested by anti-HCV; if the second testing yielded an indeterminate result, the sample was considered as negative. Seropositive individuals were submitted to a new blood sample collection to perform PCR and genotyping tests for clinical approach.

                                    HCV RNA detection and genotyping

                                    HCV RNA was detected using AMPLICOR™ (Roche Version 2.0) and extracted from 200 μL serum using the extraction reagents included in the kit, in accordance with the manufacturer’s instructions.

                                    A commercially available reverse hybridization-based line probe assay (HCV Genotype assay, LiPA™) was employed, using Amplicor’s PCR product generation. The genotypes were identified following the manufacturer’s instructions.

                                    Sample size

                                    The sample size was based on the expected prevalence of seropositivity for anti-HCV according to data from blood donors by macro-region [23]. The variance was estimated as 20% of the prevalence value for all macro-regions and 30% for the Federal District [22]. The sample size achieved in this study matched the number of individuals estimated for each region and age group as previously reported.

                                    As in the original protocol the analysis of risk factors for anti-HCV positivity was planned for the entire country, the number of seropositive individuals identified by the survey was large enough to detect differences in exposure corresponding to an odds ratio of 2, with a frequency of exposure among those not sick of 5% or more, a power of 80% and an alpha of 5%.

                                    Statistical analysis

                                    Prevalence of anti-HCV with 95% confidence intervals (95% CI) was calculated for the country, by age group and by the five macro-regions and the Federal District. The estimated number of persons infected was based on the prevalence and on the census population official data of residents from the State Capitals by age groups for the year 2000 [24]. Prevalence was adjusted for the design effect and, when necessary, for regional variations in sample fractions.

                                    The distributions of potential risk factors and their respective 95% CIs were calculated for the population aged 13 to 69 years old by macro-region. Possible associations between HCV infection and potential risk factors were assessed by calculating the Odds Ratio (OR), (95% CI) and p-values. ORs were adjusted for the design effect and age, and weighted for variations in sample fractions in the different age groups and regions. Variables that were associated with the outcome in this age-adjusted analysis (p < 0.10) were included in a multivariate regression model by means of the Generalized Linear and Latent Mixed Models (GLLAMM). The two-level analysis (individual and household) was performed using Stata 9.2 [25]. Population attributable risk—the fraction of disease in the total study population that is attributable to the exposure— was estimated for multiple factors using case–control data according to Bruzzi et al. [26]. Population attributable risk was also calculated for the ensemble of variables using an accepted procedure; it is worth noting that this estimate is not the equivalent of adding the population attributable risks for individual factors [26].

                                    A ROC curve was plotted to predict viremia for the North macro-region, and to determine the cut-off for anti-HCV positivity using the Sample rate/cut-off rate (S/CO) ratios from the individuals tested for HCV-RNA.

                                    Ethical issues

                                    Interviews and blood sample collection were conducted after written consent forms had been signed. For minors, the legal guardian’s consent was obtained. The project was approved by the National Research Ethics Committee (CONEP) of the Brazilian National Health Council and by the local Research Ethics Committees in each capital city.

                                    Each positive result was communicated by the medical doctor in charge of the field research and the negative results were sent by mail. Individuals who had a positive result were sent to referral services in each State capital or the Federal District for further clinical evaluation.

                                    Results

                                    Of 19,503 individuals aged 10 to 69 years old recruited, 2,288 were in the North region (Amazon area); 3,690 in the North-east; 3,702 in the Central-west; 1,988 in the Federal District; 3,661 in the South-east and 4,174 in the South. Of the total sample, 49.0% of the participants were aged 10–19 years old (n = 9,371) and 51.0% were 20–69 (n = 10,132). Females accounted for 54.6% of the total participants. The overall refusal rate was 2.8%, being below 1.5% for the North, Northeast and Central-west macro-regions and approximately 6% in the South and Southeast macro-regions.

                                    In Brazil, the overall weighted prevalence of HCV antibodies was 1.38% (95% CI 1.12%–1.64%) in the state capitals of the five macro-regions and the Federal District taken as a whole (Table 1). Seropositivity varied from 0.68% in the Northeast to 2.10% in the North region. In all macro-regions studied, adults (aged 20–69 years) had a higher prevalence of HCV antibodies than the younger age-group. The highest prevalence in adults was in the North region (3.22%; 95% CI 2.03%–4.41%). Prevalence peaked in those aged ≥ 60 years.
                                    Table 1

                                    Prevalence of hepatitis C antibodies and estimated population ever-infected in a representative sample of individuals living in the state capitals of Brazil, 2005–2009

                                    Setting age-group

                                    Participants

                                    Prevalence* % (95% CI)

                                    State capitals population

                                    Estimated persons ever infected n (95% CI)

                                    Brazil

                                        

                                     10 – 19

                                    9,371

                                    0.75 (0.53 – 0.98)†

                                    7,565,695

                                    56,970 (39,947 – 74,068)

                                     20 – 39

                                    5,552

                                    1.36 (1.02 – 1.71)†

                                    13,828,140

                                    188,478 (140,494 – 236,462)

                                     40 – 59

                                    3,719

                                    1.55 (1.09 – 2.01)†

                                    7,941,985

                                    122,783 (86,250 – 159,396)

                                     60 – 69

                                    891

                                    3.41 (2.03 – 4.79)†

                                    1,916,570

                                    65,432 (38,964 – 91,900)

                                    North

                                        

                                     10 – 19

                                    1,153

                                    0.99 (0.36 –1.63)

                                    860,922

                                    8,601 (3,117 – 14,093)

                                     20 – 69

                                    1,135

                                    3.22 (2.02 – 4.41)

                                    2,093,269

                                    67,403 (42,472 – 92,334)

                                    Northeast

                                        

                                     10 – 19

                                    1,840

                                    0.38 (0.10 – 0.66)

                                    2,054,539

                                    7,848 (2,096 – 13,601)

                                     20 – 69

                                    1,850

                                    0.97 (0.48 – 1.47)

                                    5,358,923

                                    52,249 (25,616 – 78,883)

                                    Midwest

                                        

                                     10 – 19

                                    1,803

                                    0.99 (0.53 – 1.47)

                                    866,536

                                    8,657 (4,601 – 12,712)

                                     20 – 69

                                    1,899

                                    1.64 (1.14 – 2.13)

                                    1,326,575

                                    21,703 (15,110 – 28,296)

                                    Federal District

                                        

                                     10 – 19

                                    980

                                    0.61 (0.13 – 1.09)

                                    387,831

                                    2,374 (524 – 4,227)

                                     20 – 69

                                    1,008

                                    1.09 (0.50 – 1.68)

                                    1,202,183

                                    13,116 (6,011 – 20,245)

                                    Southeast

                                        

                                     10 – 19

                                    1,794

                                    0.90 (0.49 – 1.31)

                                    3,295,110

                                    29,755 (16,212 – 43,298)

                                     20 – 69

                                    1,867

                                    1.63 (1.07 – 2.18)

                                    11,661,984

                                    189,857 (124,783 – 254,814)

                                    South

                                        

                                     10 – 19

                                    1,801

                                    0.51 (0.12 – 0.89)

                                    586,270

                                    2,978 (715 – 5,241)

                                     20 – 69

                                    2,373

                                    1.70 (1.08 – 2.32)

                                    2,043,761

                                    34,826 (22,175 – 47,456)

                                    * Prevalence adjusted for random effect.

                                    † Prevalence adjusted for random effect and weighted by regions.

                                    The estimated number of persons ever infected in the Brazilian State capitals and Federal District was 430,658 (95% CI 349,714–511,289), based on the HCV prevalence and the size of the population. The largest infected population was in the Southeast region (190,254; 95% CI 136,110–244,548).

                                    For the overall data set, anti-HCV positivity was similar between genders according to the two age strata (Figure 1). When stratified by macro-region, anti-HCV prevalence was similar between men and women in most of the studied settings, except in the Central-West macro-region, where men had a statistically significant higher prevalence of HCV infection than women (chi-square = 4.9, p < 0.03) (Figure 1).
                                    http://static-content.springer.com/image/art%3A10.1186%2F1471-2334-13-60/MediaObjects/12879_2012_2256_Fig1_HTML.jpg
                                    Figure 1

                                    Prevalence of hepatitis C infection in a representative sample of State capitals of the five Brazilian regions by sex and age group.

                                    Of 9,671 households investigated, ten contained two individuals testing positive for anti-HCV and one household contained three. Thirty participants were simultaneously seropositive for anti-HCV and anti-HBc antibodies.

                                    Of 236 anti-HCV positive individuals, 120 (50.8%) had their blood collected for HCV-RNA testing in all the macro-regions and Federal District, with the exception of the North macro-region, owing to operational constraints. Of those retested, 36.7% (95% IC 28.4%-45.6%) were HCV-RNA positive. The ROC curve suggested that the cut-off point of 0.4, which yielded the higher sensitivity and specificity, was OD = 4.0. Using this parameter, 32.1% individuals would also be considered infected, that is, HCV-RNA positive, in the North region. The comparison of the participants who were retested (n = 120) versus those not retested (n = 116) in relation to variables associated with HCV infection that remained in the multivariate model showed that both groups were similar in terms of age group, use of injected and sniffed drugs, use of glass syringes and hospitalization. The main difference was in socio-economic conditions (chi-square = 8.8, p = 0.012) suggesting that those better-off were more likely to be retested. Assuming the frequency of HCV-RNA-positive individuals among the estimated persons ever infected in the present study, it would be expected that there were approximately 158,000 (122,000 to 196,000) patients infected with HCV.

                                    The genotypes and subgenotypes detected were 1 (n = 3); 1a (n = 9); 1b (n = 16); 2b (n = 2); 3a (n = 14).

                                    Tables 2, 3, 4 show the age-adjusted odds ratio for the association between blood-related variables, sexual behavior and drug use and HCV infection for all the State capitals taken together. There was a positive association between anti-HCV antibodies and ever having received a blood transfusion (OR = 1.83, 95% CI 1.01–3.30), having been hospitalized in the previous 12 months (OR = 1.87, 95% CI 1.04–3.38), or having had a tattoo (OR = 1.63, 95% CI 0.95–2.80), the latter presenting a borderline p value (Table 2). Having a current partner with hepatitis was positively associated with HCV antibodies and negatively associated with partner history of STD (Table 3). Ever having used injected drugs was strongly associated with HCV antibodies (OR = 12.79, 95% CI 5.19–31.50). A statistically significant association was also found with ever having used sniffed drugs (OR = 3.50, 95% CI 1.87–6.54) and previous use of a glass syringe (OR = 1.67, 95% CI 1.09–2.56) (Table 4). As for socioeconomic variables, sewage disposal (lack of sewage system versus public system) was associated with HCV antibodies (OR = 1.90, 95% CI 1.06–3.40).
                                    Table 2

                                    Age-adjusted odds ratio of hepatitis C infection for blood route-related factors in the urban Brazilian population

                                    Risk factors

                                    Participants

                                    HCV infection n (%)

                                    ORadjusted(CI)*

                                    p-value

                                    Dental treatment

                                        

                                     Never

                                    2,034

                                    22 (1.1)

                                    1.0

                                    -

                                     Past 12 months

                                    8,655

                                    99 (1.1)

                                    0.77 (0.41 – 1.47)

                                    0.436

                                     Ever

                                    8,772

                                    114 (1.3)

                                    0.91 (0.47 – 1.77)

                                    0.795

                                    Surgery

                                        

                                     Never

                                    11,481

                                    108 (0.9)

                                    1.0

                                    -

                                     Past 12 months

                                    1,689

                                    31 (1.8)

                                    1.38 (0.72 – 2.64)

                                    0.327

                                     Ever

                                    6,270

                                    96 (1.5)

                                    0.93 (0.60 – 1.42)

                                    0.730

                                    Blood transfusion

                                        

                                     Never

                                    18,122

                                    196 (1.1)

                                    1.0

                                    -

                                     Past 12 months

                                    256

                                    10 (3.9)

                                    1.45 (0.65 – 3.21)

                                    0.365

                                     Ever

                                    843

                                    24 (2.9)

                                    1.83 (1.01 – 3.30)

                                    0.043

                                    Hospitalization

                                        

                                     Never

                                    13,095

                                    143 (1.1)

                                    1.0

                                    -

                                     Past 12 months

                                    1,308

                                    25 (1.9)

                                    1.87 (1.04 – 3.38)

                                    0.037

                                     Ever

                                    5,032

                                    67 (1.3)

                                    1.46 (0.97 – 2.19)

                                    0.065

                                    Endoscopy

                                        

                                     Never

                                    16,695

                                    190 (1.1)

                                    1.0

                                    -

                                     Past 12 months

                                    904

                                    13 (1.4)

                                    1.14 (0.53 – 2.47)

                                    0.729

                                     Ever

                                    1,785

                                    29 (1.6)

                                    1.17 (0.64 – 2.15)

                                    0.615

                                    Tatoo

                                        

                                     No

                                    18,034

                                    202 (1.1)

                                    1.0

                                    -

                                     Yes

                                    1,459

                                    33 (2.3)

                                    1.63 (0.95 – 2.80)

                                    0.076

                                    Body piercing

                                        

                                     No

                                    17,859

                                    215 (1.2)

                                    1.0

                                    -

                                     Yes

                                    1,634

                                    20 (1.2)

                                    0.97 (0.51 – 1.87)

                                    0.935

                                    Health-care related job

                                        

                                     No

                                    15,424

                                    195 (1.3)

                                    1.0

                                    -

                                     Yes

                                    1,206

                                    20 (1.7)

                                    0.91 (0.46 – 1.80)

                                    0.783

                                    * Adjusted for random effect and age weighted by age groups and regions.

                                    Table 3

                                    Age-adjusted odds ratio of hepatitis C infection for sexual behavioral-related factors in the urban Brazilian population

                                    Risk factors

                                    Participants

                                    HCV infection n (%)

                                    ORadjusted(95% CI)*

                                    p-value

                                    Initiated sexual life

                                        

                                     No

                                    4,137

                                    36 (0.9)

                                    1.0

                                    -

                                     Yes

                                    12,468

                                    179 (1.4)

                                    0.97 (0.56 – 1.68)

                                    0.912

                                    Condom use

                                        

                                     Always

                                    3,831

                                    37 (1.0)

                                    1.0

                                    -

                                     Sometimes

                                    1,944

                                    29 (1.5)

                                    1.71 (0.85 – 3.43)

                                    0.132

                                     Never

                                    6,077

                                    103 (1.7)

                                    1.04 (0.56 – 1.96)

                                    0.888

                                    Current partner had hepatitis

                                        

                                     No

                                    10,263

                                    140 (1.4)

                                    1.0

                                    -

                                     Yes

                                    445

                                    13 (2.9)

                                    2.16 (1.00 – 4.65)

                                    0.050

                                    Bisexual partner

                                        

                                     No

                                    11,417

                                    158 (1.4)

                                    1.0

                                    -

                                     Yes

                                    276

                                    4 (1.4)

                                    0.83 (0.29 – 2.42)

                                    0.737

                                    Unknown

                                    775

                                    17 (2.2)

                                    1.41 (0.69 – 2.87)

                                    0.343

                                     Sexual partners with STD

                                        

                                     No

                                    11,086

                                    164 (1.5)

                                    1.0

                                    -

                                     Yes

                                    819

                                    10 (1.2)

                                    0.43 (0.20 – 0.93)

                                    0.031

                                    Previous STD

                                        

                                     No

                                    11,079

                                    153 (1.4)

                                    1.0

                                    -

                                     Yes

                                    1,301

                                    25 (1.9)

                                    0.96 (0.50 – 1.83)

                                    0.901

                                    Another sexual partner besides current

                                        

                                     No

                                    4,703

                                    64 (1.4)

                                    1.0

                                    -

                                     Yes

                                    6,978

                                    101 (1.4)

                                    1.10 (0.73 – 1.65)

                                    0.657

                                    * Adjusted for random effect and age weighted by age groups and regions.

                                    Table 4

                                    Age-adjusted odds ratio of hepatitis C infection for drug use-related factors in the urban Brazilian population

                                    Risk factors

                                    Participants

                                    HCV infection n (%)

                                    ORadjusted(95% CI)*

                                    p-value

                                    Ever use of smoked drugs

                                        

                                     No

                                    14,726

                                    177 (1.2)

                                    1.0

                                    -

                                     Yes

                                    1,839

                                    38 (2.1)

                                    1.47 (0.88 – 2.46)

                                    0.141

                                    Ever use of inhalated drugs

                                        

                                     No

                                    16,293

                                    206 (1.3)

                                    1.0

                                    -

                                     Yes

                                    265

                                    09 (3.4)

                                    2.68 (0.85 – 8.45)

                                    0.093

                                    Ever use of sniffed drugs

                                        

                                     No

                                    15,827

                                    189 (1.2)

                                    1.0

                                    -

                                     Yes

                                    749

                                    25 (3.3)

                                    3.50 (1.87 – 6.54)

                                    0.000

                                    Ever use of injected drugs

                                        

                                     No

                                    16,463

                                    204 (1.2)

                                    1.0

                                    -

                                     Yes

                                    70

                                    11 (15.7)

                                    12.79 (5.19 – 31.5)

                                    0.000

                                    Injections with glass syringe

                                        

                                     No

                                    13,450

                                    148 (1.1)

                                    1.0

                                    -

                                     Yes

                                    2,931

                                    63 (2.2)

                                    1.67 (1.09 – 2.56)

                                    0.019

                                    Alcohol consumption

                                        

                                     None

                                    10,237

                                    129 (1.3)

                                    1.0

                                    -

                                     Light consumption

                                    5,376

                                    67 (1.2)

                                    0.75 (0.50 – 1.12)

                                    0.155

                                     Heavy consumption

                                    978

                                    16 (1.6)

                                    0.84 (0.41 – 1.69)

                                    0.619

                                     Dependent

                                    41

                                    03 (7.3)

                                    2.88 (0.79 – 10.49)

                                    0.108

                                    * Adjusted for random effect and age weighted by age groups and regions.

                                    The multivariate analysis shows that a history of injected drug use (OR = 6.65; 95% CI 2.47–17.91) and sniffed drug use (OR = 2.59; 95% CI 1.34–5.81) were strongly associated to HCV infection. Other risk factors were the use of a glass syringe (OR = 1.52; 95% CI 0.97–2.36) and ever-hospitalization, both with a borderline p value. A proxy measure of socioeconomic conditions (sewage disposal) was also associated with testing positive for anti-HCV. Age, introduced in the model as a continuous variable, showed that there was an increase in the likelihood of becoming infected of 0.02% for each year of life (OR = 1.02; 95% CI 1.01–1.04 (Table 5). The proportion of cases attributable to each variable in the final multivariate model ranged from 4.2% to 10.2%, and the proportion attributable to all factors together was 39.9%.
                                    Table 5

                                    Multivariate analysis of hepatitis C infection for potential risk factors and estimated population attributable risk in the urban Brazilian population

                                    Factors associated

                                    ORadjusted(95% CI)*

                                    p-value

                                    Population Arributable risk %

                                    Age

                                    1.02 (1.01 – 1.04)

                                    0.001

                                    -

                                    Ever use of injected drugs

                                       

                                     No

                                    1.0

                                    -

                                     

                                     Yes

                                    6.65 (2.47 – 17.91)

                                    < 0.0001

                                    4.3

                                    Ever use of sniffed drugs

                                       

                                     No

                                    1.0

                                    -

                                     

                                     Yes

                                    2.59 (1.34 – 5.01)

                                    0.005

                                    7.2

                                    Injection with glass syringe

                                       

                                     No

                                    1.0

                                    -

                                     

                                     Yes

                                    1.52 (0.97 – 2.36)

                                    0.065

                                    10.2

                                    Hospitalization

                                       

                                     Never

                                    1.0

                                    -

                                     

                                     Past 12 months

                                    1.90 (1.03 – 3.51)

                                    0.040

                                    5.0

                                     Ever

                                    1.44 (0.94 – 2.21)

                                    0.094

                                    8.7

                                    Sewage disposal

                                       

                                     Public system

                                    1.0

                                    -

                                     

                                     Septic tank

                                    1.39 (0.91 – 2.12)

                                    0.125

                                    8.8

                                     Other destination

                                    2.53 (1.38 – 4.65)

                                    0.003

                                    3.9

                                       

                                    39.9**

                                    * Adjusted for random effect and age weighted by age groups and regions.

                                    ** Summary population attributable risk.

                                    Discussion

                                    This first Brazilian nationwide hepatitis survey showed 1.38% HCV positivity in a representative sample of 19,503 adolescents and adults in all macro-regions. Approximately 1.3 million individuals would be expected to be anti-HCV-positive in the country under the assumption of similar anti-HCV antibody prevalence for the Brazilian population as a whole (population of 169.8 million, as of 2000). The current population-based survey detected a lower prevalence of anti-HCV antibody than that previously reported by the WHO for the country [2], shifting from intermediate to low prevalence.

                                    The few nationwide population-based studies have reported prevalence of anti-HCV antibodies of 1.3% in the USA [6] and 1.0% in France [27], which are similar to that found in this study. An increase in anti-HCV prevalence with age was found in the large census surveys conducted in the USA. Peak HCV prevalence was found among adults aged 30-49 years in the first census survey (1988–1994), shifting to 40–49 years in the later surveys (1999–2002; 2003–2006; 2007–2008) [6]. Although our results also showed increased HCV exposure with age, the peak prevalence was observed in the older age group. Increased HCV infection among the adult population has also been reported in selected regions of Brazil [11, 13] and among first-time blood donors [28].

                                    The highest prevalence of the HCV marker (2.1%) was in the North macro-region of Brazil, which is below the 2.5% prevalence threshold for low endemicity according to WHO criteria [2]. However, most of the estimated infected individuals resided in the Southeast region (~220,000), since approximately 14 million individuals aged 10 to 69 years live in these highly urbanized State capitals that include the cities of Sao Paulo and Rio de Janeiro. The burden of infection in absolute terms thus has an uneven geographical distribution, concentrated in the South and Southeast macro-regions. According to the official surveillance system, the residents of the southeast and south had higher HCV detection rates (above the national figures) than the residents in the north, northeast and central-west from 2003 through 2009 (unpublished data). One likely explanation for the higher detection rates in the south and southeast may be better access to and organization of the health system of these regions [29]. The surveillance data for HCV may also underestimate the prevalence of HCV infection in the country as a whole, in view of the prevalence found by this population-based survey. Under-reporting of HCV infection by other national surveillance systems has also been discussed for Europe [30, 31] and for the USA [32], pointing to the need for updated population-based studies for public health purposes.

                                    Interestingly, those participating in this national survey showed similarities in terms of the risk profile for HCV acquisition, with slightly higher frequencies for blood transfusion (7.8%), tattoos (12.2%) and sniffed drugs (5.9%) in the south macro-region (data not shown). In this household survey, injected and sniffed drug use was strongly associated with HCV antibodies in multivariate analysis. These findings accord with the literature regarding injected drug use as the main source of transmission of HCV infection in developed countries and with the reports of sniffed drugs as another important mode of HCV acquisition in developing countries, as previously reported in selected Brazilian populations [33]. In the present survey, a history of previous hospitalization and use of injections with glass syringe not related to drug use were also associated with HCV seropositivity, both of which have been widely reported as epidemiological risk factors for HCV infection worldwide [1, 2]. In our survey, a socioeconomic marker of extreme poverty (no sewage disposal) remained associated with anti-HCV prevalence (OR = 2.53), after adjusting for confounding factors. Although there is no biological plausibility for social deprivation and increased risk of HCV infection, the former may be a surrogate marker for being at risk for HCV transmission. In a countrywide survey in France, social factors remained associated with HCV infection after adjusting for known risk factors as in our study. Nevertheless, having ever used drugs, having tattoos, and having being incarcerated were correlated with social deprivation [8]. The USA HCV prevalence survey also reported a correlation of infection by drug use with low family income, high number of sex partners, and non-injected drug use [5]. Furthermore, studies using cross-sectional designs such as our survey cannot sort out the temporal relationships between the potential risk factors and the outcomes.

                                    The estimated population attributable risk indicates that the risk factors identified explain approximately 40% of all cases of infection. The higher population attributable risk percent found for “ever use of sniffed drugs” (7.2%) than for “ever use of injected drugs” (4.3%) was probably due to the lower frequency of IDUs than of sniffed drug users in the studied population. Although we cannot evaluate to what extent there was an underreporting of injection drug use because of stigma and discrimination in our study, the frequencies we found (data not shown) seem consonant with the national survey conducted by the Brazilian Center of Information on Psychotropic drugs (CEBRID) and a study among hospitalized patients [34, 35]. In relation to IDU, as the population attributable risk is related to the general population (injecting and non-injecting drug users), the attributable risk among the exposed (injecting drug users) will be greater and therefore the proportion of infection in that group that could be prevented by eliminating the exposure would be larger among IDUs.

                                    One limitation of this survey is that it included a sampling of the capital cities of the five Brazilian macro-regions and the Federal District and was not designed as part of a census survey [22] as was NHANES [3]. Therefore, the low prevalence of anti-HCV detected in all macro-regions may not reflect the levels of infection outside the study settings, such as in rural areas, and specific pockets. Since this investigation was planned as a population study, specific high-risk groups for HCV acquisition, such as institutionalized or incarcerated individuals, were excluded. Therefore, the prevalence reported here should be considered a conservative estimate of anti-HCV prevalence.

                                    It should be noted that, in view of the low frequency of HCV antibodies for the country as a whole, the risk factor analysis was performed for the entire data set and not by macro-region. This had been anticipated by the study protocol [22].

                                    Another methodological issue regarding the interpretation of our findings is that a sole marker of HCV infection may lead to misclassification, owing to false positive results. However, it would be a non-differential misclassification, underestimating the associations reported. For this reason, the strength of association may also be conservative [36]. We acknowledge that the approximately 40% of HCV-RNA positivity among anti-HCV positive individuals was lower than expected. Potential explanations for this are poor specificity of the anti-HCV antibody assay, poor sensitivity of the HCV-RNA assay, or a true biologically higher HCV-RNA spontaneous clearance rate in the Brazilian setting. In our study, high sensitivity (poor specificity) of anti-HCV was identified because of the cutoff point recommended by the manufacturer; this may have increased false positive rates. In this regard, the ROC curve showed that a cutoff point of OD =4.0 yielded higher specificity on the detection of viremia. The assay used (AMPLICOR™ Roche Version 2.0) for HCV-RNA testing detects 60 International Units per ml. A concern on the detection of hepatitis C virus RNA in a large population survey is the specimen handling and storage. Nevertheless, we have followed the laboratory procedures and storage recommendations and all tests were performed in a reference laboratory where quality control was assured. In addition, data analysis showed that the frequency of viremia found was consistent across macro-regions. In general, about 25% of infected HCV individuals have spontaneous viral clearance. A systematic review of longitudinal studies on spontaneous viral clearance showed that clearance was more frequent in females and in acute clinical hepatitis C infection [37]. Furthermore, a genome-wide association study showed that individuals with favorable IL28B genotypes are more likely to clear HCV infection than are those with unfavorable alleles, which could have implications in viral clearance ability among different ethnic groups [38]. In the South of Brazil, IL28B polymorphism was associated with spontaneous clearance of hepatitis C infection among co-infected HCV and HIV/AIDS patients [39].

                                    Although this survey covered a large geographical area and enrolled a large population sample in all State Capitals and Federal District, there was low refusal rate in all macro-regions, which was slightly higher in the wealthier regions: South and Southeast. This indicates the feasibility of nationwide-studies.

                                    In this survey, subtypes 1a, 1b, 2b and 3a were detected in the general Brazilian population within the macro-regions. Overall, genotype 1 was predominant, as reported in the literature, which suggests that genotype 1a and 1b are the most frequent HCV genotypes worldwide, accounting for 60% of the infections [2]. The geographical distribution of hepatitis C virus genotypes in Brazil among a large sample of chronic hepatitis C patients from all regions has also shown a predominance of genotype 1 followed by genotype 3 [19], in accordance with our findings. Among injecting drug-users, the circulating subtypes were 1 and 3 in the studies conducted between 1994–1997 and 1999–2001 in Rio de Janeiro [40]. Our study was not designed to identify the diversity of HCV genotypes among the regions. We are aware that not all anti-HCV positive individuals were tested for HCV-RNA, which may limit the extrapolation of these results.

                                    Conclusions

                                    The present survey measured the hepatitis C infection prevalence among the general population for the country and macro-regions, and classifies Brazil as a low-endemicity area. Nevertheless, the large estimated absolute numbers of infected individuals indicate the burden of the disease in the near future. This implies costs for the health-care system and society as a whole, as pointed out in other studies [41, 42]. This issue needs to be addressed by studies that evaluate the economic impact of HCV infection. The known risk factors explain less than 50% of the infected cases, limiting the prevention strategies. Nevertheless, our findings regarding risk behaviors associated with HCV infection showed that there is still room for improving strategies for reducing transmission among drug users. Another recommendation would be to study the extent to which nosocomial transmission is occurring in our setting. Finally, there is a need for targeting individuals living in poverty, in view of the evidence found of an association between infection and social deprivation.

                                    Abbreviations

                                    HCV: 

                                    Hepatitis C Virus

                                    NHANES: 

                                    National Health and Nutrition Examination Survey

                                    IDU: 

                                    Intravenous drug use

                                    WHO: 

                                    World Health Organization

                                    IBGE: 

                                    The Brazilian Institute of Geography and Statistics

                                    OD: 

                                    Optical density

                                    RNA: 

                                    Ribonucleic acid

                                    PCR: 

                                    Polymerase Chain Reaction

                                    OR: 

                                    Odds ratio

                                    95%CI: 

                                    95% Confidence Interval

                                    GLLAMM: 

                                    Generalized Linear and Latent Mixed Models.

                                    Declarations

                                    Acknowledgments

                                    We thank to National Hepatitis Study Group whose names are listed below, by their collaboration and support in the organization and development of the study at local level: Zulma Medeiros (Centro de Pesquisas Aggeu Magalhães-FIOCRUZ), Demócrito Miranda Filho (Universidade de Pernambuco), Maria Mabel Melo (LACEN-PE), Conceiçao Sá (Hemocentro—HEMOPI-PI), Carlos Henrique Nery Costa (Universidade Federal do Piaui), Arnaldo de Jesus Dominici (Universidade Federal do Maranhao), Maria das Graças Aragão (Universidade Federal do Maranhao), Elizabeth de Souza Lima (LACEN-MA), Jose Milton de Castro Lima (Universidade Federal do Ceara), Jose Wellington Oliveira Lima (FUNASA-CE), Maria do Carmo Vidal Gadelha Lima (LACEN-CE), Gilmar Amorim de Sousa (Universidade Federal do Rio Grande do Norte), Marcia Araujo Barreto (Universidade Federal do Rio Grande do Norte), Manoel Jaime Xavier Filho (Universidade Federal da Paraiba), Joria Viana Guerreiro (Universidade Federal da Paraiba), Dayse Mercia Cavalcanti de Oliveira (FUNASA-AL), Maria Rosileide Bezerra Alves (Secretaria Estadual de Saude de Alagoas), Ivoneide Moreira de Oliveira Barros (LACEN-PB), Tereza Virginia Silva B.Nascimento (Universidade Federal de Sergipe), Lucia Maria Sayde de Azevedo Tavares (FUNASA-SE), Raymundo Parana Ferreira Filho (Universidade Federal da Bahia), Jose Tavares-Neto (Universidade Federal da Bahia), Maria Alice Sant’Anna Zarife (LACEN-BA), Rodrigo Sebba Aires (Secretaria Estadual de Saúde de Goias), Fluvia Pereira A. da Silva (Secretaria Municipal de Saúde de Goiania), Beatriz Maranhao Bariani (LACEN-GO), Jose Ivan Aguiar (Universidade Federal do Mato Grosso do Sul), Eugenio Barros (Secretaria Municipal de Saúde-Campo Grande), Gilza Bastos dos Santos (LACEN-MS), Francisco Souto (Universidade Federal do Mato Grosso), Cor Jesus Fernando Fontes (Universidade Federal do Mato Grosso), Virgınia Correia de Azevedo (LACEN-MT), Roberto de Melo Dusi (Secretaria Estadual de Saude - DF), Lıdia Maria Pinto Luna (LACEN-DF), Jose Carlos da Fonseca (Universidade Federal do Amazonas), Leila Melo Brasil (Fundacao Medicina Tropical do Amazonas), Katia Biscuola de Campos (Programa Nacional de Prevencao e Controle das Hepatites Virais/SVS/ Ministerio da Saude), Cirley Maria de Oliveira Lobato (Serviço de Assistência Especializada - Secretaria Estadual de Saúde- AC), Mônica de Breu Moraes (Secretaria Estadual de Saúde-Acre, Programa Estadual de Hepatites), Janaina Mazaro (LACEN, Rio Branco, Acre), Maria de Nazaré Miranda de Cavalcante (Secretaria de Estado da Saúde de Amapá); Clóvis Omar Sá Miranda (Secretaria de Saúde do Estado do Amapá, Macapá, Amapa, Brazil), Márcia Socorro Cavalcante Porcy (LACEN-Amapa, Brazil), Ana Ruth Silva de Araújo (Universidade Federal do Amazonas), Marlúcia da Silva Garrido (Fundação de Vigilância em Saúde do Amazonas-Departamento de Vigilância Epidemiológica), Márcia Noronha Barbosa (Fundação de Vigilância em Saúde-AM), Raimundo Aldo Tavares (Secretaria Estadual de Saúde Pública - PA), Maria de Fátima Chaves Oliveira (Secretaria Estadual de Saúde-PA), José Américo Lozich de Aquino (LACEN-PA), Juan Miguel Villalobos-Salcedo (Universidade Federal de Rondônia), Fábio Luiz Storer (Faculdade São Lucas, Porto Velho- RO), Luiz Renerys de Lima Pinheiro (Secretaria de Saúde de Estado da Saúde de Roraima), Jacqueline de Aguiar Barros (Secretaria de Estado da Saúde de Roraima), Juliana Cristina Ferreira Lima (LACEN-RR), Nádja Duarte Oliveira Souza (Universidade Federal do Tocantins), Arlete Lopes da Cunha Otoni (Secretaria de Estado da Saúde do Tocantins); Gisele Christina O. Silva (LACEN-TO), Paraná Dominique Muzillo (Universidade Federal do Paraná), Lucimar Bozza (Secretaria Municipal de Saúde de Curitiba); Yatiyo Matsui Moriya (LACEN-Paraná), Esther Buzaglo Dantas-Correa (Universidade Federal de Santa Catarina), Ana Cristina Vidor (Prefeitura Municipal de Florianópolis), Ana Duarte Cardoso (LACEN-SC), Alexandro Alves (Universidade Federal de Ciências da Saúde de Porto Alegre), Maria Eugênia Pinto (Universidade Federal de Ciências da Saúde de Porto Alegre), Zenaida Marion Alves Nunes Lyra Vidal (Instituto de Pesquisa Biologica IPB Lacen RS), Angelica Espinosa Barbosa Miranda (Universidade Federal do Espírito Santo), Moacir Soprani (Secretaria Estadual de Saúde do Espírito Santo), Patrícia Rolim Prates (LACEN-ES), Cristiane Alves Villela-Nogueira (Universidade Federal do Rio de Janeiro), Mariza Miranda Theme-Filha (ENSP-FIOCRUZ), Carlos Augusto da Silva Fernandes (LACEN – RJ), Angela Maria Miranda Spina (Instituto Adolfo Lutz), Flair José Carrilho (Universidade de São Paulo), Antônio Alci Barone (Universidade de São Paulo), Rosângela Teixeira (Universidade Federal de Minas Gerais–UFMG); Keli Bahia Felicíssimo Zocratto (Centro Universitário Newton Paiva), Sônia Conceição Diniz (Secretaria de Estado da Saúde de Minas Gerais).

                                    We also thank to the Brazilian Ministry for Health and Pan American Health Organization by the financial support, to the University of Pernambuco Foundation by the administrative support and to the National Institute of Science and Technology for Health Technology Assessment (CNPq) by providing scholarships to R.A.A.X.(n. 308311/2009-4), C.M.T.M. (n. 306489/2010-4) and M.D.T.(n. 306928/2010-8).

                                    Authors’ Affiliations

                                    (1)
                                    Universidade de Pernambuco, Faculdade de Ciências Médicas de Pernambuco, Hospital Universitário Oswaldo Cruz
                                    (2)
                                    Instituto do Fígado de Pernambuco
                                    (3)
                                    Instituto de Patologia Tropical e Saúde Publica, Departamento de Saúde Coletiva, Universidade Federal de Goiás
                                    (4)
                                    Departamento de Medicina Tropical, Universidade Federal de Pernambuco
                                    (5)
                                    Instituto Adolfo Lutz
                                    (6)
                                    Faculdade de Ciências da Saúde, Departamento de Saúde Coletiva. DSC - Faculdade de Saúde, Universidade de Brasília
                                    (7)
                                    Fundação Universidade Federal de Ciências da Saúde de Porto Alegre
                                    (8)
                                    Departamento de Epidemiologia, Faculdade de Saúde Pública, Universidade de São Paulo
                                    (9)
                                    Fundação Oswaldo Cruz, Centro de Pesquisas Aggeu Magalhães
                                    (10)
                                    Secretaria de Saúde Pública do Estado do Pará
                                    (11)
                                    Departamento de Medicina Preventiva e Social, Universidade Federal de Pernambuco
                                    (12)
                                    Hospital de Base do DF
                                    (13)
                                    Instituto Bioestatístico –IBCT

                                    References

                                    1. Shepard CW, Finelli L, Alter MJ: Global epidemiology of hepatitis C virus infection. Lancet Infect Dis 2005,5(9):558–567.PubMedView Article
                                    2. Te HS, Jensen DM: Epidemiology of hepatitis B and C viruses: a global overview. Clin Liver Dis 2010,14(1):1–21. viiPubMedView Article
                                    3. Alter MJ, Kruszon-Moran D, Nainan OV, McQuillan GM, Gao F, Moyer LA, Kaslow RA, Margolis HS: The prevalence of hepatitis C virus infection in the United States, 1988 through 1994. N Engl J Med 1999,341(8):556–562.PubMedView Article
                                    4. Amon JJ, Garfein RS, Ahdieh-Grant L, Armstrong GL, Ouellet LJ, Latka MH, Vlahov D, Strathdee SA, Hudson SM, Kerndt P, et al.: Prevalence of hepatitis C virus infection among injection drug users in the United States, 1994–2004. Clin Infect Dis 2008,46(12):1852–1858.PubMedView Article
                                    5. Armstrong GL, Wasley A, Simard EP, McQuillan GM, Kuhnert WL, Alter MJ: The prevalence of hepatitis C virus infection in the United States, 1999 through 2002. Ann Intern Med 2006,144(10):705–714.PubMed
                                    6. McQuillan GM, Kruszon-Moran D, Denniston MM, Hirsch R: Viral Hepatitis. In NCHS Data Brief. Prevention CfDCa, vol. 27. Hyattsville, MD: National Center for Health Statistics; 2010.
                                    7. Daniels D, Grytdal S, Wasley A: Surveillance for acute viral hepatitis - United States, 2007. Morbidity and mortality weekly report Surveillance summaries (Washington, DC: 2002) 2009,58(3):1–27.
                                    8. Meffre C, Le Strat Y, Delarocque-Astagneau E, Dubois F, Antona D, Lemasson JM, Warszawski J, Steinmetz J, Coste D, Meyer JF, et al.: Prevalence of hepatitis B and hepatitis C virus infections in France in 2004: social factors are important predictors after adjusting for known risk factors. J Med Virol 2010,82(4):546–555.PubMedView Article
                                    9. Kershenobich D, Razavi HA, Sanchez-Avila JF, Bessone F, Coelho HS, Dagher L, Goncales FL, Quiroz JF, Rodriguez-Perez F, Rosado B, et al.: Trends and projections of hepatitis C virus epidemiology in Latin America. Liver Int 2011,31(Suppl 2):18–29.PubMedView Article
                                    10. Brazilian Ministry of Health: Boletim Epidemiológico das Hepatites Virais 2010. Brasília: Brazilian Ministry of Health; 2010.
                                    11. Foccacia R, da Conceição O, Sette H, Sabino E, Bassit L, Nitrini D, Lomar A, Lorenço R, Vieira de Souza F, Kiffer C, et al.: Estimated Prevalence of Viral Hepatitis in the General Population of the Municipality of São Paulo, Measured by a Serologic Survey of a Stratified, Randomized and Residence-Based Population. Braz J Infect Dis 1998,2(6):269–284.
                                    12. Zarife MA, Silva LK, Silva MB, Lopes GB, Barreto ML, Teixeira Mda G, Dourado I, Reis MG: Prevalence of hepatitis C virus infection in north-eastern Brazil: a population-based study. Trans R Soc Trop Med Hyg 2006,100(7):663–668.PubMedView Article
                                    13. Ivantes CA, Silva D, Messias-Reason I: High prevalence of hepatitis C associated with familial history of hepatitis in a small town of south Brazil: efficiency of the rapid test for epidemiological survey. Braz J Infect Dis 2010,14(5):483–488.PubMedView Article
                                    14. Carneiro MA, Martins RM, Teles SA, Silva SA, Lopes CL, Cardoso DD, Vanderborght BO, Yoshida CF: Hepatitis C prevalence and risk factors in hemodialysis patients in Central Brazil: a survey by polymerase chain reaction and serological methods. Mem Inst Oswaldo Cruz 2001,96(6):765–769.PubMedView Article
                                    15. Perone C, Del Castillo DM, Pereira GL, Carvalho Nde O, Januario JN, Teixeira R: High prevalence of genotype 1 in individuals with hepatitis C in Belo Horizonte, MG. Rev Soc Bras Med Trop 2008,41(3):238–242.PubMedView Article
                                    16. Lopes CL, Teles SA, Espirito-Santo MP, Lampe E, Rodrigues FP, Motta-Castro AR, Marinho TA, Reis NR, Silva AM, Martins RM: Prevalence, risk factors and genotypes of hepatitis C virus infection among drug users, Central-Western Brazil. Rev Saude Publica 2009,43(Suppl 1):43–50.PubMedView Article
                                    17. Nishioka Sde A, Gyorkos TW, MacLean JD: Tattoos and transfusion-transmitted disease risk: implications for the screening of blood donors in Brazil. Braz J Infect Dis 2002,6(4):172–180.PubMed
                                    18. Freitas SZ, da Cunha RV, Martins RM, Teles SA, Ibanhes ML, Motta-Castro AR: Prevalence, genotypes and risk factors associated with hepatitis C virus infection in hemodialysis patients in Campo Grande, MS, Brazil. Mem Inst Oswaldo Cruz 2008,103(4):405–408.PubMedView Article
                                    19. Campiotto SPJ, Carrilho FJ, Da Silva LC SF, Spinelli V, et al.: Geographic distribution of hepatitis C virus genotypes in Brazil. Brazilian Journal of Medicine and Biology Research 2005, 38:41–49.
                                    20. Ximenes RAA, Martelli CMT, Merchán-Hamann E, Montarroyos UR, Braga MC, de Lima MLC, Cardoso MRA, Turchi MD, Costa MA, de Alencar LCA, et al.: Multilevel analysis of hepatitis A infection in children and adolescents: a household survey in the Northeast and Central-west regions of Brazil. Int J Epidemiol 2008, 37:852–861.
                                    21. Pereira LM, Martelli CM, Merchan-Hamann E, Montarroyos UR, Braga MC, de Lima ML, Cardoso MR, Turchi MD, Costa MA, de Alencar LC, et al.: Population-based multicentric survey of hepatitis B infection and risk factor differences among three regions in Brazil. AmJTrop Med Hyg 2009,81(2):240–247.
                                    22. Ximenes RA, Pereira LM, Martelli CM, Merchan-Hamann E, Stein AT, Figueiredo GM, Braga MC, Montarroyos UR, Brasil LM, Turchi MD, et al.: Methodology of a nationwide cross-sectional survey of prevalence and epidemiological patterns of hepatitis A, B and C infection in Brazil. Cad Saude Publica 2010,26(9):1693–1704.PubMedView Article
                                    23. Fonseca JC: Epidemiologia da infecção pelo vírus da hepatite C no Brasil. Gastroenterologia Endoscopia Digestiva 1999,18(supl 1):S3-S8.
                                    24. The Brazilian Institute of Geography and Statistics (IBGE): Demographic Census 2000: characteristics of the population and households, results from the population universe. Available from: http://​www.​ibge.​gov.​br/​home/​estatistica/​populacao/​censo2000/​default.​shtm. Available from: .
                                    25. Rabe-Hesketh S, Skrondal A: Multilevel and longitudinal modeling using Stata. College Station, Texas: Stata Press; 2005.
                                    26. Bruzzi P, Green SB, Byar DP, Brinton LA, Schairer C: Estimating the population attributable risk for multiple risk factors using case–control data. Am J Epidemiol 1985,122(5):904–914.PubMed
                                    27. Delarocque-Astagneau E, Meffre C, Dubois F, Pioche C, Le Strat Y, Roudot-Thoraval F, Hillon P, Silvain C, Dhumeaux D, Desenclos JC: The impact of the prevention programme of hepatitis C over more than a decade: the French experience. J Viral Hepat 2010,17(6):435–443.PubMedView Article
                                    28. Nascimento MC, Mayaud P, Sabino EC, Torres KL, Franceschi S: Prevalence of hepatitis B and C serological markers among first-time blood donors in Brazil: a multi-center serosurvey. J Med Virol 2008,80(1):53–57.PubMedView Article
                                    29. Victora CG, Barreto ML, do Carmo Leal M, Monteiro CA, Schmidt MI, Paim J, Bastos FI, Almeida C, Bahia L, Travassos C, et al.: Health conditions and health-policy innovations in Brazil: the way forward. Lancet 2011,377(9782):2042–2053.PubMedView Article
                                    30. Rantala M, van de Laar MJ: Surveillance and epidemiology of hepatitis B and C in Europe - a review. Euro Surveill 2008,13(21):pii=18880. Available online: http://​www.​eurosurveillance​.​org/​ViewArticle.​aspx?​ArticleId=​18880.
                                    31. Irving WL, Salmon D, Boucher C, Hoepelman IM: Acute hepatitis C virus infection. Euro Surveill 2008,13(21):pii=18879. Available online: http://​www.​eurosurveillance​.​org/​ViewArticle.​aspx?​ArticleId=​18879.
                                    32. Alter MJ: Epidemiology of hepatitis C virus infection. World J Gastroenterol 2007,13(17):2436–2441.PubMed
                                    33. Duailibi LB, Ribeiro M, Ronaldo L: Profile of cocaine and crack users in Brazil. Cad Saude Publica 2008,24(Supl 4):S545-S557.PubMedView Article
                                    34. Fonseca AM: Comparison between two household surveys on psychotropic drug use in Brazil: 2001 and 2004. Ciencia e Saude Coletiva 2010,15(3):663–670.PubMedView Article
                                    35. Ferreira Filho OF, Turchi MD, Laranjeira R, Castelo A: Perfil sociodemografico e de padrões de uso entre dependentes de cocaína hospitalizados. Ver Saude Publica 2003,37(6):751–759.View Article
                                    36. Schlesselmann JJ: Case–control studies. Design, conduct, analysis. Oxford: Oxford University Press; 1982.
                                    37. Micallef JM, Kaldor JM, Dore GJ: Spontaneous viral clearance following acute hepatitis C infection: a systematic review of longitudinal studies. J Viral Hepat 2006,13(1):34–41.PubMedView Article
                                    38. Thomas DL, Thio CL, Martin MP, Qi Y, Ge D, O'Huigin C, Kidd J, Kidd K, Khakoo SI, Alexander G, et al.: Genetic variation in IL28B and spontaneous clearance of hepatitis C virus. Nature 2009,461(7265):798–801.PubMedView Article
                                    39. Lunge VR, da Rocha DB, Béria JU, Tietzmann DC, Stein AT, Simon D: IL28B polymorphism associated with spontaneous clearance of hepatitis C infection in a Southern Brazilian HIV type 1 population. AIDS Res Hum Retroviruses 2012,28(2):215–219.PubMedView Article
                                    40. Oliveira ML, Bastos FI, Telles PR, Hacker Mde A, Oliveira SA, Miguel JC, Yoshida CF: Epidemiological and genetic analyses of Hepatitis C virus transmission among young/short- and long-term injecting drug users from Rio de Janeiro, Brazil. J Clin Virol 2009,44(3):200–206.PubMedView Article
                                    41. Wong JB, McQuillan GM, McHutchison JG, Poynard T: Estimating future hepatitis C morbidity, mortality, and costs in the United States. Am J Public Health 2000,90(10):1562–1569.PubMedView Article
                                    42. King LA, Le Strat Y, Meffre C, Delarocque-Astagneau E, Desenclos JC: Assessment and proposal of a new combination of screening criteria for hepatitis C in France. Eur J Public Health 2009,19(5):527–533.PubMedView Article
                                    43. Pre-publication history

                                      1. The pre-publication history for this paper can be accessed here:http://​www.​biomedcentral.​com/​1471-2334/​13/​60/​prepub

                                    Copyright

                                    © Pereira et al.; licensee BioMed Central Ltd. 2013

                                    This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://​creativecommons.​org/​licenses/​by/​2.​0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.