Table 1 Characteristics of studies included in the systematic review of detection of HBV-DNA and update of the systematic review on HCV-RNA from DBS samples compared to venous blood sampling (plasma/serum)
Author, Country, Year | Journal | Study pop and sample size | Storage conditions | DBS collection method | Plasma method PCR | DBS method PCR | Limit of detection | Specificity | Sensitivity | Correlation/Association | Effect of storage conditions | Comments |
|---|---|---|---|---|---|---|---|---|---|---|---|---|
Alidjinou Congo 2014 | Diagnostic Microbiology and Infectious Disease | 32 HBV patients | DBS at room temperature, Frozen plasma samples at −80 °C | 30 μl of plasma onto filter paper | COBAS Taqman/COBAS AmpliPrep | COBAS Taqman/COBAS AmpliPrep | Detection limit for plasma was 12 IU/ml, In 3 patients, viral load in plasma was 152, 250, and 1727 IU/mL, respectively, whereas HBV-DNA could not be quantified in DPS, but was detected | NR | 25/26 96% | Spearman correlation coefficient r = 0.84 | NR | NR |
Alhusseini Egypt 2012 | Americal Journal of Biochemistry and Biotechnology | 50 HBs Ag pos, but only 42 later included in accuracy calculation10 negative controls | Stored at −80 °C | 50 μl from whole blood sample on Watman filter paper | In house | In house | No cut off suggested, | 10/10 100% | 42/42 100% | Good correlation (r = 0.88) between DBS and plasma viral load | NR | NR |
Durgadevi India 2012 | Conference abstract | 60 hospital patients, 30 Hep B surface marker positive 22 of which were HBV-DNA pos by serum ELISA and 30 negative | DBS were stored at 25° for 4 and 7 days | NR | NR | NR | NR | NR | 22/22100% | NR | All 22 serum positive cases could be detected from DBS after 4 and after 7 days at 25° | NR |
Gupta India 1992 | J Clin Microbiology | 60 mothers with chronic HBV infection, 5 known negative laboratory staff | −20C for filter paper | 5 μl whole blood from finger prick on Whatman paper | In house (endpoint) | In house (endpoint) | Limit of detection 10E4 virus particles in each 5 μl blood spot | 13/15 86% 5/5 lab staff | 43/45 96% | NR | Complete stability was maintained at 37 °C for 5 months, less denaturated protein when stored at 37 °C | No diagnostic accuracy calculation but can be calculated from data |
Gruner 2015 Germany | Journal of Visualized Experiments | Inpatients, 299 | Temperature: −20C, 4C or ambient temperature Time: Up to 14 days | Venipuncture, 100 μl whole blood on paper card | Not specified | Not specified | Unclear, but 7 samples not detected with DBS had HBV DNA concentrations between 409 to 3643 IU/ml | NR | 93% 93/100 | NR | NR | NR |
Jardi Spain 2004 | Hepatology | 82 patients with chronic HBV infection (23 HBeAgpos, 39 HBeAgneg, 20 HBeAg inactive, 15 HBe neg under lamivudine therapy | Storage at −20 °C | 20 μl of capillary blood on 5 mm paper disks (Scheicher) | In house | In house | LOD 100cop/ml among eight samples with serum HBV DNA between 10E3 and 10E4 copies/mL, seven tested positive using DBS samples, and among four samples with detectable serum HBV-DNA levels _10E3 copies/mL, none were positive using DBS samples. | NR | 72/77 94% | Regression coefficient HBV-DNA concentration in DBS versus serum samples r(2) = 0.96 (P < .001). | Stability of unknown number of DBS samples assessed by leaving samples for several days in different conditions, no effect on HBV-DNA levels | NR |
Lira Mexico 2009 | Virology Journal | 47 HBV HBsAg positive patients | Plasma samples at −70 °C, filter paper at −20 °C | 50 μl per card (Schleicher + Schull) | QIAamp® Ultrasens® Virus kit (QIAGEN GMBH, Germany), | NR | NR | NR | 47/47100% | The Pearson correlation 0.93 (p = 0.01) | No adverse effect by sample storage from ten patients at 4 degree, 25 degree, and 37 degree C for 7 days | NR |
Mohamed France 2013 | PLOS One | 50 HBV positive patient, 10 HBV negative patients | Dried for 18 h in room temperature, then processed | 50 μl on 12 mm discs (Whatman) | Cobas AmpliPrep/Cobas Taqman HBV test, | Cobas AmpliPrep/Cobas Taqman HBV test, | limit of detection of HBV-DNA 20 IU/ml plasma, limit of detection DBS 914 IU/ml | 49/50 98% | 50/50100% | Correlation good between DBS and plasma (r2 = 0.86, p < 0.001), | No difference in HBV-DNA levels at 1,3,7 and 14 days of storage at room temperature | Lower sensitivity in patients with 1000 IU/ml |
Mossner Denmark 2016 | Cohort | 404 Prospective patients from hepatitis clinic and blood donors | Temperature: Room temperature Time 1–5 days | Finger prick, 75 μ on Whatman filter paper | HBV DNA (Ultrio Elite dHBV). COBAS® AmpliPrep/COBAS® TaqMan® 48 system (Roche Diagnostics, Basel, Switzerland) | HBV DNA (Ultrio Elite dHBV). COBAS® AmpliPrep/COBAS® TaqMan® 48 system (Roche Diagnostics, Basel, Switzerland) | Limit of detection for HBV-DNA 200 IU/ml | 7/7 100% | 53/76 70,7% | NR | Variation of 24 h to up to 7 d found no difference in stability of samples | |
Ross Germany 2013 | Virology | 150 samples | Dried overnight at room temperature, then processed | 100 μl applied to filter paper (Whatman, Schleicher + Schüll) | artus HBV LC PCR (Qiagen, Hilden, Germany) | artus HBV LC PCR (Qiagen, Hilden, Germany) | Limit of detection 100 iU/ml in plasma, 7 samples with HBV concentrations of 409–3643 IU/ml missed | 50/50100% | 93/100 93% | NR | NR | NR |
Stene Johansen Ethiopia 2016 | PLOS One | Known HBs pos patients, recruited at hospital | Dried overnight, then stored at ambient temperatures for a median of 24 days | Venipuncture, 80 μl applied to Whatman filter card | Abbott RealTime HBV assay). | Abbott RealTime HBV assay). | NR | NR | 21/21 100% (cut off <2.7 log IU/ml) 23/26 88.5% (for all plasma detectable HBV DNA) | R = 0.92 | NR | NR |
Vinikoor Zambia 2015 | Clinical Journal of Virology | 68 HBs pos patients, | Dried for 12 hours at room temperature, stored at -80C | 50 μl applied to Whatman filter paper | Cobas AmpliPrep/Cobas Taqman HBV test | Cobas AmpliPrep/Cobas Taqman HBV test | The probability of a undetectable DBS resultat a plasma viral load of 200 IU/ml was 13.8% (95% CI: 7.7–23.7) but this dropped to 1.8% (95% CI: 0.5–6.6) when a 2000 IU/ml cut-off was used and 0.2% (95% CI: 0.03–1.7) at 20,000 IU/ml. | 62/68 91% | NR | NR | NR | NR |
HCV-RNA – additional studies from upgrade of existing systematic review [18] | ||||||||||||
Dokubo US 2014 | Journal of Clinical Virology | 148 participants in a prospective study of HCV | DBS air-dried for 2 hours, then sent to another insttute, then stored at −70 °C | Fingerstick on Whatman 903 cards 0.5 ml blood | Standard diagnostics HCV TMA (Norvatis®) | Standard diagnostics HCV TMA (Norvatis®) | 100% 84/84 | 90% 43/48 | Kappa = 0.92 | NR | ||
Gruner 2015 Germany | Journal of Visualized Experiments | Inpatients, 299 | Temperature: −20C, 4C or ambient temperature Time: Up to 14 days | Venipuncture, 100 μl whole blood on paper card | Not specified | Not specified | NR | NR | 100% | NR | NR | NR |
Marins, 2017 US | Journal of Virological Methods | 48 patients with chronic HCV infection | Left to dry in ambient temperatures overnight, then in zip bags at ambient temperatures for 4 weeks | Venipuncture, 35 μl whole blood on Whatman card | COBAS® AmpliPrep/COBAS® Taqman® HCV Quantitative Test v2.0 | COBAS® AmpliPrep/COBAS® Taqman® HCV Quantitative Test | Not detected at 4.75 log10 IU/mL. | NR | 47/48 (98%) | |||
Marques Brazil 2016 | Journal of Clinical Virology | 99 (59 anti HCV/HCV RNA pos, 40 neg samples) | NR | Venipuncture, 3–5 drops on Whatman filter paper | In house | In house | 58.5 copies/ml | 100% | 35/44 (65.9%) | Kappa: 0.648 | Low variation during 3 days at ambient temperatures | NR |
Mossner Denmark 2016 | World Journal of Gastroenterology | 404 Prospective patients from hepatitis clinic and blood donors | Temperature: Room temperature Time 1–5 days | Finger prick, 75 μ on Whatman filter paper | HCV RNA (Ultrio Elite dHBV). COBAS® AmpliPrep/COBAS® TaqMan® 48 system (Roche Diagnostics, Basel, Switzerland) | HCV RNA (Ultrio Elite dHBV). COBAS® AmpliPrep/COBAS® TaqMan® 48 system (Roche Diagnostics, Basel, Switzerland) | Limit of detection for HCV-RNA 100 IU/ml | 25/26 96% | 81/84 96.4% | NR | Variation of 24 h to up to 7 d found no difference in stability of samples | NR |
Soulier, France 2016 | The Journal of Infectious Diseases | 511 patients recruited, with known serostatus for HCV | Temperature:-80 Time: | Venipuncture, 50 μl on Whatman filter paper | 1) Cobas AmpliPrep automated extractor. The Cobas TaqMan 96 analyzer 2) m2000SP automated extractor | 1) Cobas AmpliPrep automated extractor. The Cobas TaqMan 96 analyzer 2) m2000SP automated extractor | NR | 1) 100% 196/196 2)100% 196/196 | 1) 97,1% 306/315 2) 98,1% 308/314 | NR | 25 dB samples stored at ambient temperatures (24 °C) for a mean duration (±SD) of 19 ± 1 months | |