Detection of plasmid DNA from all Chlamydia trachomatis serovars with a two-step polymerase chain reaction

D. Welch, C. H. Lee, S. H. Larsen

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

A polymerase chain reaction was used to amplify a 137-base-pair sequence of DNA from a Chlamydia trachomatis plasmid. Various parameters of the polymerase chain reaction were explored, and it was found that two short steps per reaction cycle were sufficient to achieve 1012-fold amplification in less than 1 h. By use of this procedure, 10-18 g of a sequence of plasmid DNA, representing the amount of that sequence found in one C. trachomatis bacterium, was amplified to the point where it was clearly visible on an ethidium bromidestained polyacrylamide gel under UV light. DNA from intact cells from each of the 15 serovars of C. trachomatis could also be amplified for visualization. With this procedure, the presence or absence of C. trachomatis DNA in a sample could be established in less than 1.5 h. The speed and extreme sensitivity of this detection procedure may make it a useful method for the detection of C. trachomatis, and similar techniques should be possible for any type of bacteria.

Original languageEnglish (US)
Pages (from-to)2494-2498
Number of pages5
JournalApplied and environmental microbiology
Volume56
Issue number8
DOIs
StatePublished - 1990

ASJC Scopus subject areas

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

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