Detection of a heat-labile enterotoxin gene in enterotoxigenicEscherichia coli by densitometric evaluation using highly specific enzyme-linked oligonucleotide probes

  • S. Tamatsukuri
  • K. Yamamoto
  • S. Shibata
  • F. Leaño
  • T. Honda
  • T. Miwatani
Article

Abstract

Two alkaline phosphatase-conjugated 24-mer oligonucleotide probes were developed to detect the heat-labile enterotoxin gene in enterotoxigenicEscherichia coli. Probes were antisense codon sequences, which are transcribed into mRNA, of the heat-labile enterotoxin gene of enterotoxigenicEscherichia coli of human origin. Using dot-blot hybridization, probes were tested with 100 clinical isolates and evaluated by a reflectance-type densitometer. Results agreed very well with those of an immunological test, the Biken test, and a32P-labelled recombinant DNA probe. The oligonucleotide probes did not react with nucleic acids prepared from other diarrhoeagenic bacterial pathogens. Thus, the alkaline phosphatase-conjugated oligonucleotide probes seem to be highly sensitive and specific for detection of heat-labile enterotoxin-producing enterotoxigenicEscherichia coli. Moreover, the results indicate a potential usefulness for densitometric evaluation of DNA hybridization.

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Copyright information

© Friedr. Vieweg & Sohn Verlagsgesellschaft mbH 1991

Authors and Affiliations

  • S. Tamatsukuri
    • 1
    • 2
  • K. Yamamoto
    • 1
  • S. Shibata
    • 2
  • F. Leaño
    • 1
  • T. Honda
    • 1
  • T. Miwatani
    • 1
  1. 1.Department of Bacteriology and Serology, Research Institute for Microbial DiseasesOsaka UniversitySuita, OsakaJapan
  2. 2.Toyobo Research CenterOhtsu, ShigaJapan

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