title: Genotyping of Vibrio alginolyticus isolates from Daya Bay by infrequent-restriction-site PCR and pulsed-field gel electrophoresis.
abstract: Vibrio alginolyticus is a serious bacterial pathogen that hampered the whole industry of fish farming in Guangdong, China. In order to facilitate epidemiologic studies and improve the control of disease, we developed a highly efficient method of genotyping for V. alginolyticus using infrequent-restriction-site PCR (IRS-PCR) technology. With the enzyme combination of NotI-HhaI, optimized, unique, and easy-to-interpret patterns were generated. Forty-five V. alginolyticus isolates from aquatic animals and the marine environment in Daya Bay in Guangdong were fingerprinted by IRS-PCR and pulsed-field gel electrophoresis (PFGE). The reproducibility of the IRS-PCR method was high (100%) in conformity with PFGE. When primer PN-T or PN-G was used, IRS-PCR obtained the identical clustering as PFGE, producing 24 different patterns, respectively. Moreover, IRS-PCR presented a high discriminatory power (D=0.953) identical to that of PFGE. In conclusion, the IRS-PCR fingerprinting method using NotI-HhaI can provide a potentially useful tool for epidemiologic investigations of V. alginolyticus isolates.
authors.affiliation: 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, Guangzhou, PR China.
authors.forename: Chunhua
authors.initials: C
authors.lastname: Ren
authors.forename: Chaoqun
authors.initials: C
authors.lastname: Hu
authors.forename: Peng
authors.initials: P
authors.lastname: Luo
authors.forename: Chang
authors.initials: C
authors.lastname: Chen
authors.forename: Xiao
authors.initials: X
authors.lastname: Jiang
authors.forename: Qingbai
authors.initials: Q
authors.lastname: Wang
mesh_terms: D002681:China; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods*; D005838:Genotype; D016133:Polymerase Chain Reaction / Q000379:methods*; D012623:Seawater / Q000382:microbiology*; D044149:Vibrio alginolyticus / Q000235:genetics* / Q000302:isolation & purification