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5 ' nuclease assays for the loci CCR5-+/Delta 32, CCR2-V64I, and SDFI-G80IA related to pathogenesis of AIDS

  1. Author:
    Yuan, C. C.
    Peterson, R. J.
    Wang, C. D.
    Goodsaid, F.
    Waters, D. J.

  2. Author Address

    Waters DJ NCI, Frederick Canc Res & Dev Ctr, Lab Gen Divers Frederick, MD 21702 USA NCI, Frederick Canc Res & Dev Ctr, Lab Gen Divers Frederick, MD 21702 USA NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick, AIDS Vaccine Program Frederick, MD 21702 USA PE Appl Biosyst Foster City, CA 94404 USA
    1. Year: 2000
  2. Journal: Clinical Chemistry
    1. 46
    2. 1
    3. Pages: 24-30
  4. Type of Article: Article
  1. Abstract:

    Background: Variations within the human genome play important roles in human disease. To study variations related to susceptibility to AIDS, we have developed 5' nuclease assays that eliminate post-PCR molecular biology steps. Methods: TaqMan assays based on the 5' nuclease activity of Tag polymerase and fluorescent resonance energy transfer were developed to score alleles at the biallelic loci CCR5-+/Delta 32, CCR2-V641 and SDF1-G801A. For each assay, 72 samples were analyzed. Data collection and analysis were performed on the Prism 7700 Sequence Detection System. For comparison with gel electrophoresis methods, each locus was also scored on a subset of 24 samples, using restriction enzymes or single-strand conformational polymorphism (SSCP). Results: Clear allelic discrimination was obtained on each of the 72 samples for all three TaqMan assays. The TaqMan scores for the subset of 24 samples were concordant with the restriction enzyme and SSCP scores. Conclusions: Because of its simplicity, speed, and potential for automation and miniaturization, TaqMan is an excellent candidate for investigation of genetic variation in clinical, research, and forensic settings. (C) 2000 American Association for Clinical Chemistry. [References: 28]

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