Sensitive and rapid identification of Vibrio vulnificus by loop-mediated isothermal amplification.
Vibrio vulnificus is a serious bacterial pathogen for humans and aquatic animals. We developed a rapid, sensitive and specific identification method for V. vulnificus using loop-mediated isothermal amplification (LAMP) technique. A set of primers, composed of two outer primers and two inner primers, was designed based on the cytolysin gene sequence of V. vulnificus. The LAMP reaction was processed in a heat block at 65 degrees C for 60 min. The amplification products were detected by visual inspection using SYBR Green I, as well as by electrophoresis on agarose gels. Our results showed that the LAMP reaction was highly specific to V. vulnificus. This method was 10-fold more sensitive than conventional PCR. In conclusion, the LAMP assay was extremely rapid, simple, cost-effective, sensitive and specific for the rapid identification of V. vulnificus.
Ren
Chun-Hua
CH
The Key Laboratory of Applied Marine Biology of Guangdong Province and Chinese Academy of Sciences, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou, PR China.
Hu
Chao-Qun
CQ
Luo
Peng
P
Wang
Qing-Bai
QB
D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000191:economics / Q000379:methods*; D054353:Perforin / Q000235:genetics; D012680:Sensitivity and Specificity; D041261:Vibrio vulnificus / Q000235:genetics / Q000302:isolation & purification*
D001426:Bacterial Proteins; D017931:DNA Primers; D054353:Perforin