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Comparative Study
. 2008 Dec;46(12):3941-5.
doi: 10.1128/JCM.00754-08. Epub 2008 Oct 15.

Performance of a novel human immunodeficiency virus (HIV) type 1 total nucleic acid-based real-time PCR assay using whole blood and dried blood spots for diagnosis of HIV in infants

Wendy Stevens  1 , Linda Erasmus, Matsidisho Moloi, Thabo Taleng, Somaya Sarang
Affiliations
Comparative Study

Performance of a novel human immunodeficiency virus (HIV) type 1 total nucleic acid-based real-time PCR assay using whole blood and dried blood spots for diagnosis of HIV in infants

Wendy Stevens et al. J Clin Microbiol. 2008 Dec.

Abstract

The new Cobas AmpliPrep/Cobas TaqMan HIV-1 Qual test offers advanced automation for the detection of human immunodeficiency virus type 1 (HIV-1) RNA and DNA in dried blood spots (DBS) and whole blood. An analytical evaluation using an HIV-1 secondary standard yielded limits of detection of 514, 710, and 1,090 HIV RNA copies/ml for EDTA plasma, whole blood, and DBS, respectively. The precision and reproducibility of HIV-1 detection was equivalent for DBS and whole blood. Inclusivity was demonstrated for a reference panel of HIV-1 subtypes A to N. A clinical evaluation of the Cobas AmpliPrep/Cobas TaqMan HIV-1 Qual test was performed at a center for routine diagnostics in Johannesburg, South Africa, using 1,013 clinical specimens from HIV-1 exposed children. The Amplicor HIV-1 DNA test v1.5 with the MagNApure DNA isolation procedure was used as the reference method. A total of 995 valid results for whole blood with both methods yielded 691 and 303 concordant negative and positive results for the Cobas AmpliPrep/Cobas TaqMan HIV-1 Qual test, respectively. For the 800 valid DBS specimen results, 495 and 300 concordant negative and positive results were obtained, respectively. The resulting clinical specificities and sensitivities of the new test were 100% and 99.7% for whole blood and DBS, respectively. The new test was characterized by its robustness, enhanced automation, and improved sample throughput. The Cobas AmpliPrep/Cobas TaqMan HIV-1 Qual test will support early, reliable diagnosis of HIV in children in routine laboratory settings.

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Figures

FIG. 1.
FIG. 1.
Workflow of the CAP/CTM HIV-1 Qual test for preextraction processing of the three different specimen types—whole blood, DBS, and EDTA plasma—to support advanced automation and high sample throughput.
FIG. 2.
FIG. 2.
Comparison of the CAP/CTM HIV-1 Qual test target CT values for DBS and whole blood from 300 matched positive clinical specimens subjected to Bland Altman analysis. The CT value is defined as the number of PCR cycles needed to achieve a specified target threshold fluorescence signal required for reliable detection.
See this image and copyright information in PMC

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