Dried Blood Spot Sampling for Hepatitis B Virus Serology and Molecular Testing

Mohamed, Sofiane; Raimondo, Audrey; Pénaranda, Guillaume; Camus, Claire; Ouzan, Denis; Ravet, Sophie; Bourlière, Marc; Khiri, Hacène; Dukan, Patrick; Olive, Daniel; Halfon, Philippe

doi:10.1371/journal.pone.0061077

Cited by 57 publications

(67 citation statements)

References 38 publications

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“…After diluting samples to lower levels of viremia, they estimated a limit of detection in DBS of 914.1 IU/ml (standard deviation, 157.8). 7 Similar to our results, Mohamed and others reported that DBS samples had consistently lower HBV viral loads compared to plasma, with those differences ranging from 0.65 to 1.0 log IU/ml. 6,7 The average difference in viral load between sample types in our study (1.59 log IU/ml) was not unexpected since DBS have smaller input volumes of 75–100 μL of whole blood (approximately half of which is comprised of plasma) versus 650 μL used in plasma testing.…”

Section: Discussionsupporting

confidence: 92%

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Hepatitis B viral load in dried blood spots: A validation study in Zambia

Vinikoor

Zürcher

Musukuma

et al. 2015

Journal of Clinical Virology

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Background Access to hepatitis B viral load (VL) testing is poor in sub-Saharan Africa (SSA) due to economic and logistical reasons. Objectives To demonstrate the feasibility of testing dried blood spots (DBS) for hepatitis B virus (HBV) VL in a laboratory in Lusaka, Zambia, and to compare HBV VLs between DBS and plasma samples. Study design Paired plasma and DBS samples from HIV-HBV co-infected Zambian adults were analyzed for HBV VL using the COBAS AmpliPrep/COBAS TaqMan HBV test (Version 2.0) and for genotype by direct sequencing. We used Bland-Altman analysis to compare VLs between sample types and by HBV genotype. Logistic regression analysis was conducted to assess the probability of an undetectable DBS result by plasma VL. Results Among 68 participants, median age was 34 years, 61.8% were men, and median plasma HBV VL was 3.98 log IU/ml (interquartile range, 2.04–5.95). Among sequenced viruses, 28 were genotype A1 and 27 were genotype E. Bland-Altman plots suggested strong agreement between DBS and plasma VLs. DBS VLs were on average 1.59 log IU/ml lower compared to plasma with 95% limits of agreement of −2.40 to −0.83 log IU/ml. At a plasma VL ≥2,000 IU/ml, the probability of an undetectable DBS result was 1.8% (95% CI: 0.5–6.6). At plasma VL ≥20,000 IU/ml this probability reduced to 0.2% (95% CI: 0.03–1.7). Conclusions In a Zambian laboratory, we observed strong agreement between DBS and plasma VLs and high sensitivity in DBS at plasma VL ≥2,000 IU/ml. As HBV treatment expands, DBS could increase access to HBV VL testing in SSA settings.

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Section: Discussionsupporting

confidence: 92%

“…Suboptimal elution of nucleic acids from the filter paper is another explanation for the difference between DBS and plasma viral loads. In this study, we used manual elution methods but other studies 7 have utilized automated platforms to optimize the procedure.…”

Section: Discussionmentioning

confidence: 99%

Hepatitis B viral load in dried blood spots: A validation study in Zambia

Vinikoor

Zürcher

Musukuma

et al. 2015

Journal of Clinical Virology

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“…Several studies have shown decreased sensitivity of DBS compared to plasma in samples with lower viral loads[20,28,29]. In our study, we estimated that the LLD was below 500 copies/mL for HIV-RNA, 100 IU/mL for HCV-RNA and 200 IU/mL for HBV-DNA by DBS testing.…”

Section: Discussionmentioning

confidence: 73%

Dried blood spots, valid screening for viral hepatitis and human immunodeficiency virus in real-life

Mössner¹,

Staugaard²,

Jensen³

et al. 2016

WJG

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AIMTo detect chronic hepatitis B (CHB), chronic hepatitis C (CHC) and human immunodeficiency virus (HIV) infections in dried blood spot (DBS) and compare these samples to venous blood sampling in real-life.METHODSWe included prospective patients with known viral infections from drug treatment centers, a prison and outpatient clinics and included blood donors as negative controls. Five drops of finger capillary blood were spotted on filter paper, and a venous blood sample was obtained. The samples were analyzed for HBsAg, anti-HBc, anti-HBs, anti-HCV, and anti-HIV levels as well as subjected to a combined nucleic acid test (NAT) for HBV DNA, HCV RNA and HIV RNA.RESULTSSamples from 404 subjects were screened (85 CHB, 116 CHC, 114 HIV and 99 blood donors). DBS had a sensitivity of > 96% and a specificity of > 98% for the detection of all three infections. NAT testing did not improve sensitivity, but correctly classified 95% of the anti-HCV-positive patients with chronic and past infections. Anti-HBc and anti-HBS showed low sensitivity in DBS (68% and 42%).CONCLUSIONDBS sampling, combined with an automated analysis system, is a feasible screening method to diagnose chronic viral hepatitis and HIV infections outside of the health care system.

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“…Previous studies have shown that DBS can be used to reliably measure human immunodeficiency virus (HIV) RNA viral load [6–8] and hepatitis C virus (HCV) RNA viral load [9–11]. However, only a few smaller studies, and none in low-income countries, have assessed the performance of DBS for HBV DNA quantification and applied it in patient management [12–15]. …”

Section: Introductionmentioning

confidence: 99%

Dry Blood Spots a Reliable Method for Measurement of Hepatitis B Viral Load in Resource-Limited Settings

et al. 2016

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Background & AimsHepatitis B virus (HBV) quantification is essential in the management of chronic hepatitis B, both to determine treatment eligibility and in the monitoring of treatment effect. This test, however, is rarely available in resource-limited settings due to high costs and stringent requirements for shipment and storage of plasma. Dried Blood Spots (DBS) can be a convenient alternative to plasma, but its use for HBV monitoring has not been investigated under real-life conditions in Africa.MethodsThe performance of DBS in HBV quantification was investigated using a modified commercial test (Abbott RealTime HBV assay). Paired DBS and plasma samples were collected from an HBV positive cohort in Addis Ababa, Ethiopia. DBS were stored at ambient temperature for 4–39 days before shipment to the laboratory.ResultsTwenty-six paired samples were selected covering the total range of quantification, from 2.14 log IU/ml to >7 log IU/ml. HBV was detected in 21 of 21 (100%) DBS from patients with a corresponding plasma viral load above 2.70 log IU/ml. The mean difference between plasma and DBS was 0.59 log IU/ml, and the correlation was strong (R2 = 0.92). In stability studies there was no significant change in DBS viral load after storage at room temperature for up to 12 weeks.ConclusionsThis study suggests that DBS can be a feasible and reliable alternative to plasma for quantification of HBV in resource-limited settings. DBS can expand access to antiviral treatment for patients in low- and middle-income countries.

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Dried Blood Spot Sampling for Hepatitis B Virus Serology and Molecular Testing

Cited by 57 publications

References 38 publications

Hepatitis B viral load in dried blood spots: A validation study in Zambia

Hepatitis B viral load in dried blood spots: A validation study in Zambia

Dried blood spots, valid screening for viral hepatitis and human immunodeficiency virus in real-life

Dry Blood Spots a Reliable Method for Measurement of Hepatitis B Viral Load in Resource-Limited Settings

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