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Genotypic detection of enterotoxigenic Escherichia coli colonisation factors
Author(s) -
Grewal Harleen M. S.,
Sommerfelt Halvor
Publication year - 2001
Publication title -
apmis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.909
H-Index - 88
eISSN - 1600-0463
pISSN - 0903-4641
DOI - 10.1034/j.1600-0463.2001.090607.x
Subject(s) - enterotoxigenic escherichia coli , colonisation , microbiology and biotechnology , escherichia coli , biology , plasmid , fimbria , genotype , antigen , virology , dna , genetics , gene , enterotoxin , colonization
We have developed a nonradioactive colony hybridisation assay for the detection of enterotoxigenic Escherichia coli (ETEC) that harbor the structural genes for CFA/I, CS1, CS2, CS4, CS17, or PCFO166. Thus, a polynucleotide probe derived from the colonisation factor antigen I (CFA/I) operon hybridised under very low stringency conditions to total DNA from CFA/I‐producing (CFA/I), coli‐surface antigen 1 and 3 (CS1 CS3‐), CS2 CS3‐, CS4 CS6‐, CS17‐, and putative colonisation factor O166 (PCFO166)–producing enterotoxigenic Escherichia coli (ETEC). The probe did not hybridise to DNA from CS3, CFA/III CS6, CS5 CS6, CS6, CS7, or PCFO159 ETEC. Visual registration of colour intensity could be used to differentiate between CFA/I, CS4 and PCFO166‐positive strains on the one hand and strains with the genetic potential to express CS1, CS2, or CS17 on the other. As a confirmatory test, restriction fragment patterns obtained from Sau 3AI‐digested ETEC plasmid DNA could be used to distinguish between CFA/I, CS1, CS4, CS17, and PCFO166 ETEC in nonradioactive Southern blot hybridisation. The simultaneous genotypic detection of several ETEC colonisation factors will prove useful in vaccine‐oriented studies of ETEC disease.