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

详细信息    查看全文

• 作者：Ning Tang ; Shihai Huang ; John Salituro ; Wai-Bing Mak ; Gavin Cloherty ; Julie Johanson ; Yu Hong Li ; George Schneider ; John Robinson ; John Hackett Jr. ; Priscilla Swanson ; Klara Abravaya
• 关键词：Monitoring HIV-1 viral load ; Real-time PCR
• 刊名：Journal of Virological Methods
• 出版年：2007
• 出版时间：December 2007
• 年：2007
• 卷：146
• 期：1-2
• 页码：236-245
• 全文大小：447 K

文摘

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™ 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™ 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.