Reference - Detail
| RRC ID | 15628 |
|---|---|
| Author | Yamazaki W, Seto K, Taguchi M, Ishibashi M, Inoue K. |
| Title | Sensitive and rapid detection of cholera toxin-producing Vibrio cholerae using a loop-mediated isothermal amplification. |
| Journal | BMC Microbiol |
| Abstract |
BACKGROUND:Vibrio cholerae is widely acknowledged as one of the most important waterborne pathogen causing gastrointestinal disorders. Cholera toxin (CT) is a major virulence determinant of V. cholerae. Detection of CT-producing V. cholerae using conventional culture-, biochemical- and immunological-based assays is time-consuming and laborious, requiring more than three days. Thus, we developed a novel and highly specific loop-mediated isothermal amplification (LAMP) assay for the sensitive and rapid detection of cholera toxin (CT)-producing Vibrio cholerae. RESULTS:The assay provided markedly more sensitive and rapid detection of CT-producing V. cholerae strains than conventional biochemical and PCR assays. The assay correctly identified 34 CT-producing V. cholerae strains, but did not detect 13 CT non-producing V. cholerae and 53 non-V. cholerae strains. Sensitivity of the LAMP assay for direct detection of CT-producing V. cholerae in spiked human feces was 7.8 x 102 CFU per g (1.4 CFU per reaction). The sensitivity of the LAMP assay was 10-fold more sensitive than that of the conventional PCR assay. The LAMP assay for detection of CT-producing V. cholerae required less than 35 min with a single colony on thiosulfate citrate bile salt sucrose (TCBS) agar and 70 min with human feces from the beginning of DNA extraction to final determination. CONCLUSION:The LAMP assay is a sensitive, rapid and simple tool for the detection of CT-producing V. cholerae and will be useful in facilitating the early diagnosis of human V. cholerae infection. |
| Volume | 8 |
| Pages | 94 |
| Published | 2008-6-12 |
| DOI | 10.1186/1471-2180-8-94 |
| PII | 1471-2180-8-94 |
| PMID | 18547441 |
| PMC | PMC2446398 |
| MeSH | Bacterial Typing Techniques Base Sequence Cholera / microbiology* Cholera Toxin / genetics* Cholera Toxin / metabolism Feces / microbiology Humans Molecular Sequence Data Polymerase Chain Reaction / methods* Sensitivity and Specificity Vibrio cholerae / genetics Vibrio cholerae / isolation & purification* Vibrio cholerae / metabolism |
| IF | 2.989 |
| Times Cited | 90 |
| WOS Category | MICROBIOLOGY |
| Resource | |
| General Microbes | JCM 2529 JCM2530 |