Solid phase capture method for the specific amplification of microbial nucleic acids-avoidance of false-positive and false-negative reactions

Robert H. Yolken, Ana Maria Sierra-Honigmann, Raphael P. Viscidi

Research output: Contribution to journalArticle

Abstract

DNA amplification assays such as the polymerase chain reaction are being developed for the amplification of small quantities of microbial nucleic acids. These assays offer the potential for a great deal of sensitivity. However, the high level of sensitivity increases the likelihood of cross-contamination of amplified products and the generation of false-positive reactions. In addition, substances in body fluids can inhibit the efficient performance of the amplification reactions. We have developed an assay format in which microbial nucleic acids are specifically bound to a solid phase surface. Contaminating DNA and enzyme inhibitors present in the sample are removed by washing prior to the performance of the amplification reaction. We could use this system to amplify and detect small amounts of HIV DNA diluted in whole blood. The assay system could distinguish target DNA from contaminating DNA fragments generated by prior amplification reactions.

Original languageEnglish (US)
Pages (from-to)151-156
Number of pages6
JournalMolecular and Cellular Probes
Volume5
Issue number2
DOIs
StatePublished - Apr 1991

Keywords

  • DNA amplification
  • Nucleic acids
  • PCR
  • enzyme immunoassay

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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