[Research on detection of Vibrio comma and Vibro parahaemolyticus from foodstuffs using MPCR methods].
OBJECTIVE
To establish a set of multi-PCR (MPCR) methods to detect Vibrio comma O1 serogroup (EVC) and O139 serogroup, and Vibrio parahaemolyticus rapidly and sensitively from foodstuffs.


METHODS
Using T139 (specific gene of O139 serogroup), ctxB and tcpA genes from V. comma, and tdh gene from V. parahaemolyticus as target sequences, we detected the anticipative amplified bands, whose sizes were relatively 417bp, 564bp, 471bp and 202bp.


RESULTS
Excellent specificity of the amplified products could be found from both standard and wild strains of EVC, O139VC and V. parahaemolyticus. It also means that no amplified band was detected from total 35 strains of other bacilli, including salmonella, comma bacillus which do not belong to O1 and O139 serogroups. The detection limits of artificial contaminated samples such as tilapia flesh, oyster and mixture of tilapia intestines and gills were proved to be 22cfu/g in EVC, 50cfu/g in O139 and 65cfu/g in V. parahaemolyticus. Besides, it took only 8-10 hours to finish the whole process.


CONCLUSIONS
Experiment results show that MPCR is a sensitive, convenient and time saving method suitable for detection.
Xu
Long-Yan
LY
Food Inspection Laboratory of Guangzhou Entry-Exit Inspection & Quarantine Bureau, Guangzhou 510623, China.
Zhou
Hong-Bin
HB
Gao
Dong-Wei
DW
Yang
Jing
J
D000818:Animals; D005399:Fishes / Q000382:microbiology; D005506:Food Contamination / Q000032:analysis*; D010125:Ostreidae / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D017747:Seafood / Q000382:microbiology*; D012703:Serotyping; D014734:Vibrio cholerae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D014736:Vibrio parahaemolyticus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*