Rapid and specific identification of 5 human pathogenic Vibrio species by multiplex polymerase chain reaction targeted to dnaJ gene.
A multiplex polymerase chain reaction (PCR) method, specifically designed for application in routine diagnostic laboratories, was developed for identifying 5 human pathogen Vibrio species: Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio mimicus, and Vibrio alginolyticus. This assay directed toward the dnaJ gene was tested on a total of 355 strains representing 13 Vibrio species and 17 non-Vibrio species. Specific PCR fragments were produced in isolates belonging to the 5 target species and were absent from all strains other than these 5 species and non-Vibrio strains, indicating a high specificity of this multiplex PCR. The multiplex PCR for the detection of Vibrio pathogens in clinical specimens was experimentally applied to spiked stool samples. Only 1 specific amplicon was observed, corresponding to the pathogen spiked into the stool sample. The detection limitation was 10(5) to 10(6) cells per milliliter stool. Our data showed that this method represented a robust tool for the specific and rapid detection of the 5 major pathogenic Vibrio species.
Nhung
Pham Hong
PH
Department of Microbiology, Regeneration and Advanced Medical Science, Gifu University Graduate School of Medicine, Gifu 501-1194, Japan. phnhung1976@yahoo.com
Ohkusu
Kiyofumi
K
Miyasaka
Jiro
J
Sun
Xiao Song
XS
Ezaki
Takayuki
T
D015373:Bacterial Typing Techniques; D004403:Dysentery / Q000145:classification / Q000235:genetics / Q000382:microbiology; D005243:Feces / Q000382:microbiology; D050956:HSP40 Heat-Shock Proteins / Q000145:classification / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D014733:Vibrio / Q000145:classification / Q000235:genetics; D014735:Vibrio Infections / Q000145:classification* / Q000235:genetics
D050956:HSP40 Heat-Shock Proteins