Open AccessSpecies‐specific PCR detection of the fish pathogen, Vibrio anguillarum , using the amiB gene, which encodes N ‐acetylmuramoyl‐ l ‐alanine amidase
Author(s) -
Hong GyeongEun,
Kim DongGyun,
Bae JuYoon,
Ahn SunHee,
Bai Sungchul C.,
Kong InSoo
Publication year - 2007
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2006.00618.x
Subject(s) - vibrio anguillarum , microbiology and biotechnology , biology , pathogen , peptidoglycan , polymerase chain reaction , vibrio , flounder , bacteria , gene , fish <actinopterygii> , genetics , fishery
Vibrio anguillarum is the causative agent of the fish disease vibriosis and is the most intensely studied species of Vibrio . In the present study, specific primers and a PCR assay were designed to detect V. anguillarum . The primers were designed to amplify a 429‐bp internal region of the V. anguillarum amiB gene, which encodes the peptidoglycan hydrolase N ‐acetylmuramoyl‐ l ‐alanine amidase. PCR specificity was demonstrated by successful amplification of DNA from V. anguillarum and by the absence of a PCR product from 25 other Vibrio strains and various enteric bacteria. The PCR produced a 429‐bp amplified fragment from as little as 1 pg of V. anguillarum DNA. The limit of detection for this PCR technique was c . 20 bacterial colonies in 25 mg of infected flounder tissue. These results suggest that this PCR system is a sensitive and species‐specific detection method, and is possible to use as a diagnostic tool to detect V. anguillarum .