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Direct Detection by PCR of Escherichia coli O157 and Enteropathogens in Patients with Bloody Diarrhea
Author(s) -
Takeshi Kouichi,
Ikeda Tetsuya,
Kubo Akiko,
Fujinaga Yukako,
Makino Souichi,
Oguma Keiji,
Isogai Emiko,
Yoshida Shinichi,
Sunagawa Hiroyuki,
Ohyama Tohru,
Kimura Hiroo
Publication year - 1997
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1997.tb01934.x
Subject(s) - microbiology and biotechnology , biology , vibrio parahaemolyticus , shigella , campylobacter jejuni , serotype , diarrhea , polymerase chain reaction , escherichia coli , shiga toxin , stx2 , salmonella , campylobacter , bloody diarrhea , toxin , pathogen , virology , bacteria , gene , medicine , biochemistry , genetics
Direct detection of Escherichia coli O157 and foodborne pathogens associated with bloody diarrhea were achieved using polymerase chain reaction (PCR) after the preparation of DNA from stool specimens using the microspin technique. PCR was compared with cultivation and toxin production tests with respect to the efficiency of detection of each pathogen; E. coli O157, Vibrio parahaemolyticus, Salmonella serovar Enteritidis and Campylobacter jejuni. Detection of some or all of the above pathogens in clinical stool specimens was achieved using PCR. The minimum number of cells required for the detection of the above pathogens by PCR was 10 1 CFUs/0.5 g of stool sample. PCR was completed within 6 hr. The above pathogens were also detected in cultivation and toxin production tests. Partial purification of the template DNA using the microspin technique was essential for the elimination of PCR inhibitors from the DNA samples. This PCR method is an accurate, easy‐to‐read screening method for the detection of Shiga‐like toxin producing E. coli O157 and enteropathogens associated with bloody diarrhea in stool specimens.