A simple competitive RT-PCR assay for quantitation of HIV-1 subtype B and non-B RNA in plasma

Makiko Hamatake, Masako Nishizawa, Naoki Yamamoto, Shingo Kato, Wataru Sugiura

研究成果: Article査読

6 被引用数 (Scopus)

抄録

An easy, inexpensive competitive RT-PCR assay for HIV-1 RNA quantitation was constructed. A 138-bp sequence in the HIV-1 gag p24 region was selected as the target and co-amplified with competitor RNA containing an internal 44-bp deletion. Quantitation of serial dilutions of control RNA samples prepared from the LAI isolate demonstrated a good linearity (R2 = 0.991) within the range between 10 and 250 copies/sample. The detection limit of the assay was determined to be 3.8 copies/sample by Probit analysis and corresponded to 110 copies/ml in plasma. The intra-assay CV value was 9.1%, and the inter-assay value was 25.9%. Both were comparable to those obtained with commercially available HIV-1 RNA quantitation kits. The correlation efficient for the results obtained in 47 plasma samples from HIV-1-infected individuals (subtype A in 1, subtype B in 25, subtype C in 4, subtype F in 1, and CRF01_AE in 16) with the competitive RT-PCR and Cobas Amplicor HIV-1 Monitor test v1.5 was 0.956 for subtype B and 0.947 for subtype non-B. The assay devised is a good alternative for monitoring antiretroviral therapy in resource-poor countries.

本文言語English
ページ(範囲)113-117
ページ数5
ジャーナルJournal of Virological Methods
142
1-2
DOI
出版ステータスPublished - 2007 6

ASJC Scopus subject areas

  • ウイルス学

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