Fultz T, Todd J, Hamren S, Sheridan P, Collins M, Yeghiazarian T, Kern D, Chernoff D, Urdea M; National Conference on Human Retroviruses and Related Infections.
Program Abstr Second Natl Conf Hum Retrovir Relat Infect Natl Conf Hum Retrovir Relat Infect 2nd 1995 Wash DC. 1995 Jan 29-Feb 2; 93.
Chiron Corporation, Emeryville, CA.
Quantitation of virion-associated plasma RNA may be useful in assessing efficacy of antiviral therapy and may have prognostic value in determining disease progression in HIV- infected individuals. We have developed an improved method to quantitate plasma HIV RNA levels using branched DNA (bDNA) signal amplification technology. HIV RNA is concentrated from 1 ml of plasma using centrifugation, extracted with proteinase k and detergents, and then quantitated directly through a series of solid-phase probe hybridization. Target RNA amplification is not used in this assay. Enhanced sensitivity was obtained through probe design modifications that significantly reduced non-specific hybridizations that cause background signal. Simultaneously, the signal has been further amplified through improved alkaline phosphatase activity and increased bDNA binding to target probes. The combined improved technologies lead to a 10-20 fold increased molecular sensitivity. A quantitation limit of 300-1,000 HIV RNA equivalents/ml, with a dynamic linear quantitation range of 3.5 logs without specimen dilution, has been obtained. The enhanced assay sensitivity may be useful for assessing the effect of new antiviral agents on plasma viremia particularly in HIV-infected subjects with near normal CD4 counts (ie, greater than 400 cells/cc).
Publication Types:
Keywords:
- Acquired Immunodeficiency Syndrome
- Antiviral Agents
- CD4 Lymphocyte Count
- DNA
- DNA, Viral
- Evaluation Studies
- HIV
- HIV Core Protein p24
- HIV Infections
- HIV Seropositivity
- Nucleic Acid Amplification Techniques
- Nucleic Acid Hybridization
- RNA
- RNA, Viral
- Sensitivity and Specificity
- Viremia
- immunology
Other ID:
UI: 102213194
From Meeting Abstracts