A RealTime HIV-1 viral load assay for automated quantitation of HIV-1 RNA in genetically diverse group M subtypes A-H, group O and group N samples

被引：73

作者：

Tang, Ning ^{[1
]}

Huang, Shihai ^{[1
]}

Salituro, John ^{[1
]}

Mak, Wai-Bing ^{[1
]}

Cloherty, Gavin ^{[1
]}

Johanson, Julie ^{[1
]}

Li, Yu Hong ^{[1
]}

Schneider, George ^{[1
]}

Robinson, John ^{[1
]}

Hackett, John, Jr. ^{[2
]}

Swanson, Priscilla ^{[2
]}

Abravaya, Klara ^{[1
]}

机构：

[1] Abbott Mol Inc, Des Plaines, IL 60018 USA

[2] Abbott Diagnost, Abbott Pk, IL 60064 USA

来源：

JOURNAL OF VIROLOGICAL METHODS | 2007年 / 146卷 / 1-2期

关键词：

monitoring HIV-1 viral load; real-time PCR;

D O I：

10.1016/j.jviromet.2007.07.003

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The Abbott RealTime HIV-1 assay is an automated test for monitoring HIV-1 viral load in plasma samples. The assay uses reverse transcription polymerase chain reaction (RT-PCR) technology with homogeneous real-time fluorescent detection. Automated sample preparation is performed on the m2000sp (TM) instrument where RNA is isolated using magnetic microparticle technology and dispensed to a PCR tray together with the amplification reagents. The PCR tray is then transferred to the Abbott m2000rt (TM) instrument for amplification and real-time detection. The assay utilizes two distinct sets of primers and probes for HIV-1 and for internal control (IC). The IC is processed along with each sample to control for sample recovery and inhibition. The HIV-1 primer and probe sequences are targeted to the integrase (IN) region of the polymerase (pol) gene. Due to the selection of a highly conserved target region and a novel, mismatch tolerant probe design, the assay can quantitate HIV-1 group M subtypes A-H, group O, and group N isolates. The assay provides high reproducibility and a wide dynamic range, allowing quantitation. from 40 copies to 10 million copies of HIV-1 RNA per milliliter of plasma. HIV-1 RNA concentrations detected with 95% probability were 25 copies/mL with 1.0 mL of plasma, 39 copies/mL with 0.6 mL of plasma, 65 copies/mL with 0.5 mL of plasma, and 119 copies/mL with 0.2 mL of plasma. (c) 2007 Elsevier B.V. All rights reserved.

引用

页码：236 / 245

页数：10

共 53 条

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