Dileanis JA, Kunstman K, Wolinsky S, Palumbo P, Huang D, Brown R; International Conference on AIDS.

Int Conf AIDS. 2000 Jul 9-14; 13: abstract no. TuPeA3008.

J.A. Dileanis, PE Biosystems, 850 Lincoln Centre Drive, Foster City, CA 94404, United States, Tel.: +1 650 638 5718, Fax: +1 650 638 6177, E-mail: dileanjl@pebio.com

Objective: PE Biosystems has developed a version 2 HIV Genotyping System. Clade B clinical samples were analyzed to assess performance of this modified version of the PE Biosystems HIV-1 Genotyping System at three external sites. Looseness=1 Methods: Plasma samples (0.5 mL) were tested using the PE Biosystems version 2 HIV-1 Genotyping System. In the version 2 system, RT and PCR primers were modified from the currently available version 1 system, RNA isolation reagents were improved to enhance RNA stability, and the UNG contamination control system was implemented. The version 2 RT-PCR reaction yields a PCR product which is 1.8Kb in length. The Sequencing module in the version 2 kit utilizes the PE Biosystems BigDye' terminator sequencing chemistry; it permits double coverage sequencing of the entire protease gene and codons 1-315 of the reverse transcriptase gene. Sequencing reactions were subsequently analyzed on either the ABI Prism "377 DNA Sequencer or ABI Prism" 310 Genetic Analyzer. Viral loads for these samples ranged from 260 to greater than 750,000 copies/mL. Results: With the version 2 system, 111 of 119 samples (93.3%) with a viral load greater than 2000 copies per mL produced sufficient PCR product for sequencing. At viral loads between 1000 and 2000 copies per mL, 5 of 7 samples (71%) were positive. At viral loads less than 1000 copies per mL, 6 of 12 samples (50%) were positive. Sequencing data were successfully utilized in the HGS software v2.1 to determine resistance mutations in the samples. Conclusion: The version 2 HIV-1 Genotyping System provides a sensitive and reliable assay for clade B HIV-1 analysis using 0.5 mL of clinical plasma samples. The version 2 HIV-1 Genotyping System is available for Research Use Only.

Publication Types:

• Meeting Abstracts

Keywords:

• AIDS Vaccines
• Acquired Immunodeficiency Syndrome
• HIV Infections
• HIV Seropositivity
• HIV-1
• Laboratory Techniques and Procedures
• Polymerase Chain Reaction
• Proto-Oncogene Proteins c-kit
• Research Design
• Viral Load
• immunology
• reverse transcriptase, Human immunodeficiency virus 1

Other ID:

• GWAIDS0001363

UI: 102238854

From Meeting Abstracts