title: Peptide nucleic acid fluorescence in-situ hybridization for identification of Vibrio spp. in aquatic products and environments.
abstract: A peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) method was developed for specific detection of the Vibrio genus. In silico analysis by BLAST and ProbeCheck showed that the designed PNA probe targeting the 16S rRNAs was suitable for specific identification of Vibrio. Specificity and sensitivity of the probe Vib-16S-1 were experimentally verified by its reactivity against 18 strains of 9 Vibrio species and 14 non-Vibrio strains of 14 representative species. The PNA-FISH assay was able to identify 47 Vibrio positive samples from selectively enriched cultures of 510 samples of aquatic products and environments, comparable with the results obtained by biochemical identification and real-time PCR. We conclude that PNA-FISH can be an alternative method for rapid identification of Vibrio species in a broad spectrum of seafood or related samples.
authors.affiliation: Institute of Preventive Veterinary Medicine & Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, People's Republic of China; Food Safety Key Laboratory of Zhejiang Province, Zhejiang Entry-Exit Inspection and Quarantine Bureau, 126 Fuchun Road, Hangzhou 310012, People's Republic of China.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Food Safety Key Laboratory of Zhejiang Province, Zhejiang Entry-Exit Inspection and Quarantine Bureau, 126 Fuchun Road, Hangzhou 310012, People's Republic of China.
authors.forename: Ke
authors.initials: K
authors.lastname: Li
authors.affiliation: Food Safety Key Laboratory of Zhejiang Province, Zhejiang Entry-Exit Inspection and Quarantine Bureau, 126 Fuchun Road, Hangzhou 310012, People's Republic of China.
authors.forename: Shan
authors.initials: S
authors.lastname: Wu
authors.affiliation: Food Safety Key Laboratory of Zhejiang Province, Zhejiang Entry-Exit Inspection and Quarantine Bureau, 126 Fuchun Road, Hangzhou 310012, People's Republic of China.
authors.forename: Jiangbing
authors.initials: J
authors.lastname: Shuai
authors.affiliation: Institute of Preventive Veterinary Medicine & Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, People's Republic of China. Electronic address: whfang@zju.edu.cn.
authors.forename: Weihuan
authors.initials: W
authors.lastname: Fang
mesh_terms: D015373:Bacterial Typing Techniques / Q000379:methods* / Q000592:standards; D004783:Environmental Microbiology; D005516:Food Microbiology; D017404:In Situ Hybridization, Fluorescence; D020135:Peptide Nucleic Acids / Q000737:chemistry*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014733:Vibrio / Q000235:genetics / Q000502:physiology*; D014871:Water Microbiology
chemical_list: D020135:Peptide Nucleic Acids; D012336:RNA, Ribosomal, 16S
keywords: Fluorescent in situ hybridization; Peptide nucleic acid; Vibrio species