O Chityothin

Mahidol University

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P Echeverria

Centro Uruguayo de Imagenología Molecular

Wanpen Chaicumpa

Siriraj Hospital

J Seriwatana

Walter Reed Army Institute of Research

U Vongsthongsri

Mahidol University

C Tirapat

Mahidol University

David N. Taylor

Texas Chiropractic College

U Patamaroj

S Tharavanij

Mahidol University

S L Moseley

University of Washington

Orntipa Sethabutr

Armed Forces Research Institute of Medical Science

Cooperative Institutions

Mahidol University

Armed Forces Research Institute of Medical Science

University of Maryland, Baltimore

Mahidol Oxford Tropical Medicine Research Unit

Walter Reed Army Institute of Research

National Public Health Laboratory

Stanford University

Centro Uruguayo de Imagenología Molecular

Naval Medical Research Command

Armed Forces Institute of Pathology

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Detection of enterotoxigenic Escherichia coli in water by filter hybridization with three enterotoxin gene probes

Journal of Clinical Microbiology (1982)

P Echeverria J Seriwatana O Chityothin Wanpen Chaicumpa C Tirapat

The DNA hybridization assay for genes encoding for Escherichia coli enterotoxins was used to examine water specimens in Thailand. In a reconstruction experiment, the DNA hybridization assay was 10(4) times more sensitive than testing random E. coli in the Y-1 adrenal and suckling mouse assays in identifying enterotoxigenic E. coli (ETEC) in water. Drinking and bathing water collected from 2 of 10 different homes of individuals with ETEC-associated diarrhea and 6% (5 of 78) and 11% (11 of 78) of drinking and bathing water samples collected from homes of individuals with diarrhea without ETEC infections, as well as 6% (5 of 77) and 8% (6 of 77) of drinking and bathing water collected from homes in which no inhabitants had diarrhea, were homologous with the DNA probes. Ten E. coli from each of the 31 water specimens which contained bacteria which were homologous with the DNA probes were tested in the Y-1 adrenal and suckling mouse assay. In only 2 of these 31 specimens could ETEC be identified with the standard assays. The DNA hybridization assay is a much more sensitive means of detecting organisms carrying genes coding for enterotoxin production than testing 10 individual colonies in the Y-1 adrenal and suckling mouse assays. This novel application of recombinant DNA technology provides a sensitive method of detecting organisms carrying genes coding for enterotoxin, and this method will be useful in defining the epidemiology of ETEC.

Enterotoxigenic Escherichia coli

DNA–DNA hybridization

Hybridization probe

Verocytotoxin

10.1128/jcm.16.6.1086-1090.1982

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Prevalence of heat-stable II enterotoxigenic Escherichia coli in pigs, water, and people at farms in Thailand as determined by DNA hybridization

Journal of Clinical Microbiology (1984)

P Echeverria J Seriwatana U Patamaroj S L Moseley A. McFarland

The DNA hybridization assay employing a 460-base-pair fragment of DNA encoding for the methanol-insoluble form of heat-stable toxin (ST-II) was used to determine the prevalence of ST-II enterotoxigenic Escherichia coli (ETEC) in pigs, people, and water at 57 farms in Sri Racha, Thailand. ST-II ETEC was found in 62 (3%) of 2,110 suckling, 181 (32%) of 560 weaned, and 4 (1%) of 457 adult pigs examined. Of 62 suckling pigs with ST-II ETEC infections 21% had diarrhea, but none of 185 infected older pigs had diarrhea. ST-II ETEC was found more frequently in suckling pigs with diarrhea than without diarrhea (13 of 146 versus 49 of 1,964) (P less than 0.001). ST-II ETEC was detected in water collected from 3 of 57 clay jars containing water used to bathe at three pig farms, in 1 jar used to bathe immediately after working in the barn, and from one farmer who did not have a recent history of diarrhea. Evidence of this organism was not found in 245 other individuals living on the pig farms or in 220 inhabitants and 114 water specimens collected at tapioca farms nearby. In Thailand ST-II ETEC was found in suckling pigs with diarrhea but was infrequently found in humans.

Enterotoxigenic Escherichia coli

Barn

10.1128/jcm.19.4.489-491.1984

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Detection of heat-stable enterotoxigenic Escherichia coli by hybridization with an RNA transcript probe

Journal of Clinical Microbiology (1987)

O Chityothin Orntipa Sethabutr P Echeverria David N. Taylor U Vongsthongsri

Heat-stable enterotoxigenic Escherichia coli was identified by nucleotide hybridization with RNA transcripts of the gene encoding heat-stable A-2 enterotoxin. Radiolabeled enterotoxin gene RNA transcripts are easier to prepare and avoid the preparation of cloned DNA probes that can be nonspecific if they contain cloning vector DNA.

Enterotoxigenic Escherichia coli

Heat-stable enterotoxin

Cloning (programming)

Hybridization probe

10.1128/jcm.25.8.1572-1573.1987

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