Quantification of HIV-RNA from dried blood spots using the Siemens VERSANT® HIV-1 RNA (kPCR) assay

Objectives Simplified methods for virological monitoring in resource-limited settings are increasingly needed. We evaluated the performance of the VERSANT® HIV-1 RNA (kPCR) assay for the determination of HIV-1 viral load from dried blood spots (DBS). Assay sensitivity and correlation with plasma qua...

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Bibliographic Details
Published in:Journal of antimicrobial chemotherapy 2011-12, Vol.66 (12), p.2823-2826
Main Authors: Pirillo, Maria Franca, Recordon-Pinson, Patricia, Andreotti, Mauro, Mancini, Maria Grazia, Amici, Roberta, Giuliano, Marina
Format: Article
Language:English
Subjects:
Antibiotics. Antiinfectious agents. Antiparasitic agents
Automation - methods
Biological and medical sciences
Blood - virology
Correlation analysis
Desiccation - methods
HIV
HIV Infections - virology
HIV-1 - isolation & purification
Human immunodeficiency virus
Human viral diseases
Humans
Immunodeficiencies
Immunodeficiencies. Immunoglobulinopathies
Immunopathology
Infectious diseases
Medical sciences
Molecular Diagnostic Techniques - methods
Pharmacology. Drug treatments
Plasma
Reagent Kits, Diagnostic
Ribonucleic acid
RNA
RNA, Viral - blood
Sensitivity and Specificity
Specimen Handling - methods
Viral diseases
Viral diseases of the lymphoid tissue and the blood. Aids
Viral Load
Virology
Virology - methods
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Summary:Objectives Simplified methods for virological monitoring in resource-limited settings are increasingly needed. We evaluated the performance of the VERSANT® HIV-1 RNA (kPCR) assay for the determination of HIV-1 viral load from dried blood spots (DBS). Assay sensitivity and correlation with plasma quantification values were assessed. Methods A total of 98 DBS were prepared from fresh blood samples of HIV-infected patients. DBS were kept at room temperature for 6 weeks or 7 months before processing while the corresponding plasma samples were stored at −80°C. DBS were first pre-treated in a special DBS buffer. The DBS extracts and the plasma samples were then purified and amplified using the VERSANT assay reagents. Results In the first series of tests, performed after 6 weeks of storage, there was good correlation between quantification of viral load in plasma and in DBS (r = 0.95, P 1000 copies/mL. The sensitivity and specificity of the DBS assay were 88.2% [95% confidence interval (CI) 79.4-93.6] and 69.2% (95% CI 42.0-87.4), respectively. Using the 5000 copies/mL threshold (defining virological failure in resource-limited settings), both positive and negative predictive values were high (95.2% and 87.5%, respectively). After 7 months of storage there was a modest decrease in the detection rate and less significant correlations for samples with HIV-RNA
ISSN:0305-7453
1460-2091
DOI:10.1093/jac/dkr383