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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)

From: Diagnostic accuracy of detection and quantification of HBV-DNA and HCV-RNA using dried blood spot (DBS) samples – a systematic review and meta-analysis

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