The Use of Transcription-based Amplification Systems in the Diagnosis of HIV-1 Infections

  • Deborah Y. Kwoh
  • Thomas R. Gingeras
Part of the Applied Virology Research book series (AVIR, volume 3)

Abstract

The advances in nucleic acid sequencing and detection by hybridization have opened new approaches to the diagnosis of human pathogens. While nucleic acid hybridization made detection of target sequences highly specific and/or relatively rapid in comparison to standardized immunological or culture-based assays, the methodology was slow in being adapted to clinical settings for two major reasons. First, the most sensitive nucleic acid hybridization techniques requires the use of radiolabeled probes making everyday utilization and waste management problematic. Secondly, even using radioactivity, nucleic acid hybridization is unable to detect targets below approximately 104 molecules (Chou and Merigan, 1982). This limitation in sensitivity poses a major stumbling block for the use of probes especially in viral diagnostics since several clinically important viruses such as human T-cell lymphoma/leukemia virus type 1 and 2 (HTLV-1 and 2), cytomegalovirus (CMV) and especially human immunodeficiency virus type 1 (HIV-1) frequently are present clinical samples well below the level of sensitivity achievable by most nucleic acid-based hybridization assays (Landry, 1990). A major breakthrough in the utility of nucleic acid hybridization as a diagnostic tool occurred with the introduction of in vitro nucleic acid target amplification. The development of the polymerase chain reaction (PCR) in 1985 (Saiki et al.,1985) permitted the amplification of target nucleic acid sequences over a million-fold, thereby allowing a single copy of a target molecule to be detected in a clinical sample. The only other diagnostic tool comparable in sensitivity is the in vitro propagation by culturing of the pathogen.

Keywords

Hepatitis Delta Virus Polymerase Chain Reaction Protocol Nucleic Acid Hybridization Polymerase Chain Reaction Technology Promoter Binding Site 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer Science+Business Media New York 1994

Authors and Affiliations

  • Deborah Y. Kwoh
    • 1
  • Thomas R. Gingeras
    • 1
  1. 1.Life Sciences Research LaboratoryBaxter Diagnostics, Inc.San DiegoUSA

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