Cloning and sequence of a region of Vibrio cholerae O139 Bengal and its use in PCR-based detection.
We isolated and characterized a Vibrio cholerae O139 Bengal-specific DNA region by arbitrary PCR. The fragment contains open reading frames encoding two potential glycosyltransferases possibly involved in capsular polysaccharide or lipopolysaccharide biosynthesis. In order to evaluate the possibility that this region could be used for the specific detection of V. cholerae O139 Bengal, a PCR system was established. The specificity and sensitivity of the PCR were investigated by analyzing 240 strains within the family Vibrionaceae and 178 stains of other gram-negative bacteria. All V. cholerae O139 Bengal strains tested were positive, and none of the 384 control strains were amplified. The sensitivity of the assay was 10(2) CFU/ml.
Falklind
S
S
Karolinska Institute, Department of Immunology, Microbiology, Pathology and Infectious Diseases, Huddinge University Hospital, Sweden.
Stark
M
M
Albert
M J
MJ
Uhlén
M
M
Lundeberg
J
J
Weintraub
A
A
D000595:Amino Acid Sequence; D001431:Bacteriological Techniques / Q000706:statistics & numerical data; D001459:Bangladesh / Q000453:epidemiology; D001483:Base Sequence; D002771:Cholera / Q000453:epidemiology / Q000382:microbiology; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004196:Disease Outbreaks; D005069:Evaluation Studies as Topic; D016695:Glycosyltransferases / Q000235:genetics; D006801:Humans; D017720:Molecular Epidemiology; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012475:Salmonella / Q000235:genetics; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D012703:Serotyping; D013045:Species Specificity; D014734:Vibrio cholerae / Q000145:classification / Q000201:enzymology / Q000235:genetics*
D017931:DNA Primers; D004269:DNA, Bacterial; D016695:Glycosyltransferases