Assessment of dried blood spot samples as a simple method for detection of hepatitis B virus markers

Villar, Lívia Melo; Oliveira, Jaqueline Correia de; Cruz, Helena Medina; Yoshida, Clara Fumiko Tachibana; Lampe, Elisabeth; Lewis-Ximenez, Lia Laura

doi:10.1002/jmv.22138

Cited by 72 publications

(84 citation statements)

References 24 publications

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“…Only a few reports, which directly compare different elution buffers in the context of DBS testing, are available. Villar et al 36 , e.g., recorded almost equivalent elution capacities for all buffers used, but PBS/BSA 0.5% resulted in the lowest level of non-specific reactivity. A very similar observation was made by Croom and co-workers 44 when applying the specimen diluent of the Genetics Systems rLAV EIA for the elution of anti-HCV antibodies from DBS.…”

Section: Parametersmentioning

confidence: 92%

See 1 more Smart Citation

Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques

Grüner

Stambouli

Roß

2015

JoVE

116

105

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The idea of collecting blood on a paper card and subsequently using the dried blood spots (DBS) for diagnostic purposes originated a century ago. Since then, DBS testing for decades has remained predominantly focused on the diagnosis of infectious diseases especially in resourcelimited settings or the systematic screening of newborns for inherited metabolic disorders and only recently have a variety of new and innovative DBS applications begun to emerge. For many years, pre-analytical variables were only inappropriately considered in the field of DBS testing and even today, with the exception of newborn screening, the entire pre-analytical phase, which comprises the preparation and processing of DBS for their final analysis has not been standardized. Given this background, a comprehensive step-by-step protocol, which covers al the essential phases, is proposed, i.e., collection of blood; preparation of blood spots; drying of blood spots; storage and transportation of DBS; elution of DBS, and finally analyses of DBS eluates. The effectiveness of this protocol was first evaluated with 1,762 coupled serum/DBS pairs for detecting markers of hepatitis B virus, hepatitis C virus, and human immunodeficiency virus infections on an automated analytical platform. In a second step, the protocol was utilized during a pilot study, which was conducted on active drug users in the German cities of Berlin and Essen.

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Section: Parametersmentioning

confidence: 92%

“…Storage at -20 °C or 4 °C did not result in a substantial improvement. In contrast, a study conducted thirty years later 36 tested…”

Section: Parametersmentioning

confidence: 95%

Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques

Grüner

Stambouli

Roß

2015

JoVE

116

105

View full text Add to dashboard Cite

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“…The diagnostic accuracy of HBsAg detection (ELISA) using DBS compared to the accuracy with serum or plasma has been assessed in the past and found to be very good, with sensitivity and specificity at 96 to 100% and at 97 to 100%, respectively (28)(29)(30).…”

Section: Njai Et Almentioning

confidence: 99%

Validation of Rapid Point-of-Care (POC) Tests for Detection of Hepatitis B Surface Antigen in Field and Laboratory Settings in the Gambia, Western Africa

et al. 2015

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f Hepatitis B virus (HBV) infection is a leading cause of death in sub-Saharan Africa (SSA). Point-of-care tests for hepatitis B surface antigen (HBsAg) could be an ideal tool for a large-scale HBV screening/treatment program in SSA. Using data from the PROLIFICA (Prevention of Liver Fibrosis and Cancer in Africa) program, we conducted a cross-sectional study to assess the diagnostic accuracy of three point-of-care tests (Determine, Vikia, and Espline) for the detection of HBsAg in the field or a laboratory setting in the Gambia. In the field, we used finger-prick whole blood for the Determine and Vikia tests and dried blood spots for the reference standard test (AxSYM HBsAg enzyme-linked immunosorbent assay [ELISA]). In the laboratory we used serum for the Determine, Espline, and reference test (Architect chemiluminescent microparticle immunoassay). Of 773 participants recruited at the community and 227 known chronic HBV carriers (1,000 subjects in total), 293 were positive for HBsAg. The sensitivity and specificity of the Determine test were 88.5% and 100% in the field and 95.3% and 93.3% in the laboratory setting, respectively. The sensitivity and specificity were 90.0% and 99.8% for the Vikia test (in the field) and 93.9% and 94.7% for the Espline test (in the laboratory). There was no evidence that one kit was better than another. Most of the patients with false-negative results (18/19) were classified as inactive chronic carriers. In summary, the three point-of-care tests had acceptable ranges of diagnostic accuracy. These tests may represent accurate, rapid, and inexpensive alternatives to serology testing for the screening of HBV infection at field level in SSA.

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“…As a result, DBS have been used as an alternative sampling method for laboratory diagnosis, epidemiological studies, and therapeutic monitoring of viral infections. Villar et al (19) used enzyme-linked immunosorbent assays (ELISAs) for HBsAg and anti-HBs detection in DBS samples. For HBsAg, their assay had a sensitivity of 97.62% and specificity of 96.7% and for anti-HBs, the values were 78% and 97.3%, respectively.…”

Section: Discussionmentioning

confidence: 99%

Hepatitis B virus DNA stability in plasma samples under short-term storage at 42°C

Almeida

Espírito-Santo

Sousa

et al. 2015

Braz J Med Biol Res

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We evaluated the stability of hepatitis B virus (HBV) DNA in plasma samples stored at 42°C for external quality assessment (EQA) panels of viral load. To assess the stability of plasma samples containing different concentrations of HBV DNA, serial dilutions of HBV-infected samples with a viral load of 6.40 log(10) IU/mL were made to yield viral loads of 5, 4, and 3 log(10) IU/mL. These were incubated at 42°C for up to 7 days and then frozen at -70°C. Viral load testing for HBV DNA was performed for all samples using COBAS¯ AmpliPrep/COBAS¯ TaqMan¯ HBV Test (v.2.0, Roche, Switzerland). Results were compared with fresh frozen plasma samples as a benchmark to establish acceptable measurements on the days following sample collection. Although the results of this study demonstrated a decrease in HBV DNA viral load ranging from 0.005 to 0.30 log(10) IU/mL after storage at 42°C for up to 7 days, these values did not exceed 0.5 log(10), which is the estimated intra-assay variation for molecular tests. Thus, the insignificant decrease in viral load suggests that shipment of HBV in plasma samples at temperatures of up to 42°C is permissible if they are frozen within 7 days.

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Assessment of dried blood spot samples as a simple method for detection of hepatitis B virus markers

Cited by 72 publications

References 24 publications

Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques

Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques

Validation of Rapid Point-of-Care (POC) Tests for Detection of Hepatitis B Surface Antigen in Field and Laboratory Settings in the Gambia, Western Africa

Hepatitis B virus DNA stability in plasma samples under short-term storage at 42°C

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