title: Optimization and head-to-head comparison of MISSR-PCR, ERIC-PCR, RAPD and 16S rRNA evolutionary clock for the genotyping of Vibrio cholerae isolated in China.
abstract: OBJECTIVE
  To establish a new genotyping method for Vibrio cholerae and compare it with other methods.
  
  
  METHODS
  In the current study, a modified inter simple sequence repeat-polymerase chain reaction (MISSR-PCR) system was developed via several rounds of optimisation. Comparison study was then conducted between MISSR-PCR and three other methods, including enterobacterial repetitive intergenic consensus sequences-based PCR (ERIC-PCR), randomly amplified polymorphic DNA (RAPD) and 16S rRNA evolutionary clock, for the detection and genetic tracing of Vibrio cholerae isolated from seafood in China.
  
  
  RESULTS
  The results indicated that the MISSR-PCR system could generate the highest polymorphic fingerprinting map in a single round PCR and showed the best discriminatory ability for Vibrio cholerae genotyping by clearly separating toxigenic/nontoxigenic strains, local/foreign strains, and O1/O139/non-O1/non-O139 serogroup strains, comparing to ERIC-PCR, RAPD and 16S rRNA evolutionary clock. Moreover, the MISSR-PCR is superior to previously described traditional simple sequence repeat based PCR method on genotyping by more clearly separating different clusters.
  
  
  CONCLUSIONS
  To the best of our knowledge, this is the first head-to-head comparison of four detection and genotyping methods for Vibrio cholerae The MISSR-PCR system established here could serve as a simple, quick, reliable and cost-effective tool for the genotyping and epidemiological study.
authors.forename: Q H
authors.initials: QH
authors.lastname: Mo
authors.forename: H B
authors.initials: HB
authors.lastname: Wang
authors.forename: H
authors.initials: H
authors.lastname: Tan
authors.forename: S L
authors.initials: SL
authors.lastname: An
authors.forename: Z L
authors.initials: ZL
authors.lastname: Feng
authors.forename: Q
authors.initials: Q
authors.lastname: Wang
authors.forename: J C
authors.initials: JC
authors.lastname: Lin
authors.affiliation: Key Laboratory of Diarrhea Disease Detection Zhuhai International Travel Healthcare Center, Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai, China.
authors.forename: Z
authors.initials: Z
authors.lastname: Yang
mesh_terms: D002681:China; D014644:Genetic Variation; D060005:Genotyping Techniques / Q000379:methods*; D017747:Seafood / Q000382:microbiology; D014734:Vibrio cholerae / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification