European Journal of Clinical Microbiology and Infectious Diseases

, Volume 10, Issue 12, pp 1048–1055

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

DOI: 10.1007/BF01984927

Cite this article as:
Tamatsukuri, S., Yamamoto, K., Shibata, S. et al. Eur. J. Clin. Microbiol. Infect. Dis. (1991) 10: 1048. doi:10.1007/BF01984927

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.

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