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Multiplex polymerase chain reaction to detect toxigenic Vibrio cholerae and to biotype Vibrio cholerae O1
Author(s) -
Shangkuan Y.H.,
Show Y.S.,
Wang T.M.
Publication year - 1995
Publication title -
journal of applied bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 0021-8847
DOI - 10.1111/j.1365-2672.1995.tb03136.x
Subject(s) - vibrio cholerae , amplicon , microbiology and biotechnology , multiplex polymerase chain reaction , biology , polymerase chain reaction , primer (cosmetics) , cholera toxin , toxin , multiplex , gene , bacteria , genetics , chemistry , organic chemistry
A multiplex polymerase chain reaction (PCR) was developed to identify cholera toxin‐producing Vibrio cholerae and to biotype V. cholerae O1. Enterotoxin‐producing V. cholerae strains were identified with a primer pair that amplified a fragment of the ctxA2‐B gene. Vibrio cholerae O1 strains were simultaneously differentiated into biotypes with three primers specified for the hlyA gene in the same reaction. The hlyA amplicon in the multiplex PCR serves as an internal control when testing toxin‐producing strains, as hlyA gene sequences exist in all tested V. cholerae strains. Enrichment of V. cholerae present on oysters for 6 h in alkaline peptone water before detection by a nested PCR with internal primers for ctxA2‐B gene yielded a detection limit lower than 3 colony‐forming units (cfu) per gram of food.