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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

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Development of candidate reference reagent for HIV-1 RNA and comparison analysis for different HIV-1 RNA quantitative assay

Borae G. Park1, 2 / 2 / Jee-Hye Choi2, 3 / Jina Park2, 3 / Sung Soon Kim4 / Jin-Sook Wang4 / Mee Kyung Kee4 / Ju-yeon Choi4

1Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Republic of Korea

2Department of Laboratory Medicine, College of Medicine, Chung-Ang University, Seoul, Republic of Korea

3Department of Life Science, College of Natural Science, Chung-Ang University, Seoul, Republic of Korea

4Division of AIDS, Center for Immunology and Pathology, National Institute of Health, Korea Center for Disease Control and Prevention, Seoul, Republic of Korea

Corresponding author: Ae Ja Park, MD, Department of Laboratory Medicine, College of Medicine, Chung-Ang University, 224-1, Heuksuk-Dong, Dongjak-Gu, Seoul 156-755, Republic of Korea Phone: +82 2 6299 2717, Fax: +82 2 6298 8630

Citation Information: Clinical Chemistry and Laboratory Medicine. Volume 49, Issue 9, Pages 1519–1524, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: 10.1515/CCLM.2011.231, June 2011

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Background: Human immunodeficiency virus type-1 (HIV-1) RNA viral load is a surrogate marker that is routinely used to determine indications for, and monitor the effectiveness of HIV-1 treatment. We developed three reagents for potential use in routine quality control of HIV-1 RNA quantitative assays. In this report, we compare the stability of these re-agents in storage and compare their performance in three different HIV-1 RNA quantitative assays.

Methods: The candidate reagents were derived from readily available pre-existing reagents and examined for stability at different storage temperatures. They were compared in three commercially available HIV-1 RNA quantitative assays: the Cobas TaqMan HIV-1 Test (Cobas TaqMan), the RealTime HIV-1 Assay (Abbott RealTime), and the NucliSens EasyQ HIV-1 Assay v1.1 (NucliSens EasyQ).

Results: The candidate reagent derived from an HIV culture supernatant (candidate CS) was the most stable of the three candidates and showed good reproducibility. Candidate CS yielded the highest HIV-1 titer of the three candidates in the Cobas TaqMan assay and the lowest HIV-1 titer and stability of the three candidates in the NucliSens EasyQ system.

Conclusions: The candidate CS is the most appropriate of the three candidate reagents for quantitative testing of HIV-1 RNA. This working reagent should be useful for use in routine calibration for quality control in centers with limited financial resources. The Cobas TaqMan assay tended to yield higher viral load results than the other assays when used with our three candidate reagents.

Keywords: HIV-1 RNA; quantitation; real-time PCR; stability; working reagent

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