Journal article
Performance Characteristics of the QUANTIPLEX HIV-1 RNA 3.0 Assay for Detection and Quantitation of Human Immunodeficiency Virus Type 1 RNA in Plasma
Journal of clinical microbiology, Vol.38(8), pp.2837-2845
08/2000
DOI: 10.1128/JCM.38.8.2837-2845.2000
PMCID: PMC87124
PMID: 10921936
Abstract
The QUANTIPLEX HIV-1 RNA assay, version 3.0 (a branched DNA, version 3.0, assay [bDNA 3.0 assay]), was evaluated by analyzing spiked and clinical plasma samples and was compared with the AMPLICOR HIV-1 MONITOR Ultrasensitive (ultrasensitive reverse transcription-PCR [US-RT-PCR]) method. A panel of spiked plasma samples that contained 0 to 750,000 copies of human immunodeficiency virus type 1 (HIV-1) RNA per ml was tested four times in each of four laboratories (1,344 assays). Negative results (<50 copies/ml) were obtained in 30 of 32 (94%) assays with seronegative samples, 66 of 128 (52%) assays with HIV-1 RNA at 50 copies/ml, and 5 of 128 (4%) assays with HIV-1 RNA at 100 copies/ml. The assay was linear from 100 to 500,000 copies/ml. The within-run standard deviation (SD) of the log
10
estimated HIV-1 RNA concentration was 0.08 at 1,000 to 500,000 copies/ml, increased below 1,000 copies/ml, and was 0.17 at 100 copies/ml. Between-run reproducibility at 100 to 500 copies/ml was <0.10 log
10
in most comparisons. Interlaboratory differences across runs were ≤0.10 log
10
at all concentrations examined. A subset of the panel (25 to 500 copies/ml) was also analyzed by the US-RT-PCR assay. The within-run SD varied inversely with the log
10
HIV-1 RNA concentration but was higher than the SD for the bDNA 3.0 assay at all concentrations. Log-log regression analysis indicated that the two methods produced very similar estimates at 100 to 500 copies/ml. In parallel testing of clinical specimens with low HIV-1 RNA levels, 80 plasma samples with <50 copies/ml by the US-RT-PCR assay had <50 copies/ml when they were retested by the bDNA 3.0 assay. In contrast, 11 of 78 (14%) plasma samples with <50 copies/ml by the bDNA 3.0 assay had ≥50 copies/ml when they were retested by the US-RT-PCR assay (median, 86 copies/ml; range, 50 to 217 copies/ml). Estimation of bDNA 3.0 values of <50 copies/ml by extending the standard curve of the assay showed that these samples with discrepant results had higher HIV-1 RNA levels than the samples with concordant results (median, 34 versus 17 copies/ml;
P
= 0.0051 by the Wilcoxon two-sample test). The excellent reproducibility, broad linear range, and good sensitivity of the bDNA 3.0 assay make it a very attractive method for quantitation of HIV-1 RNA levels in plasma.
Details
- Title: Subtitle
- Performance Characteristics of the QUANTIPLEX HIV-1 RNA 3.0 Assay for Detection and Quantitation of Human Immunodeficiency Virus Type 1 RNA in Plasma
- Creators
- Alejo Erice - Department of Laboratory Medicine & Pathology and Department of Medicine, Division of Infectious Diseases, University of Minnesota, Minneapolis, MinnesotaDonald Brambilla - Department of Laboratory Medicine & Pathology and Department of Medicine, Division of Infectious Diseases, University of Minnesota, Minneapolis, MinnesotaJames Bremer - Department of Laboratory Medicine & Pathology and Department of Medicine, Division of Infectious Diseases, University of Minnesota, Minneapolis, MinnesotaJ. Brooks Jackson - Department of Laboratory Medicine & Pathology and Department of Medicine, Division of Infectious Diseases, University of Minnesota, Minneapolis, MinnesotaRobert Kokka - Department of Laboratory Medicine & Pathology and Department of Medicine, Division of Infectious Diseases, University of Minnesota, Minneapolis, MinnesotaBelinda Yen-Lieberman - Department of Laboratory Medicine & Pathology and Department of Medicine, Division of Infectious Diseases, University of Minnesota, Minneapolis, MinnesotaRobert W Coombs - Department of Laboratory Medicine & Pathology and Department of Medicine, Division of Infectious Diseases, University of Minnesota, Minneapolis, Minnesota
- Resource Type
- Journal article
- Publication Details
- Journal of clinical microbiology, Vol.38(8), pp.2837-2845
- Publisher
- American Society for Microbiology
- DOI
- 10.1128/JCM.38.8.2837-2845.2000
- PMID
- 10921936
- PMCID
- PMC87124
- ISSN
- 0095-1137
- eISSN
- 1098-660X
- Language
- English
- Date published
- 08/2000
- Academic Unit
- Pathology; VPMA - Administration
- Record Identifier
- 9984047639602771
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