Genotypic detection of enterotoxigenic Escherichia coli colonisation factors.
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 Sau3AI-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.
Grewal
H M
HM
Department of Microbiology and Immunology, The Gade Institute, University of Bergen and Haukeland Hospital, Norway. Harleen.Grewal@cih.uib.no
Sommerfelt
H
H
D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics*; D015342:DNA Probes; D004269:DNA, Bacterial / Q000235:genetics; D004768:Enterotoxins / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000276:immunology / Q000472:pathogenicity; D036781:Fimbriae Proteins; D005798:Genes, Bacterial; D005838:Genotype; D006801:Humans; D009693:Nucleic Acid Hybridization; D010957:Plasmids / Q000235:genetics
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D015342:DNA Probes; D004269:DNA, Bacterial; D004768:Enterotoxins; C022924:colonization factor antigens; D036781:Fimbriae Proteins