Peptide nucleic acid fluorescence in-situ hybridization for identification of Vibrio spp. in aquatic products and environments

Xiaofeng Zhang; Ke Li; Shan Wu; Jiangbing Shuai; Weihuan Fang

doi:10.1016/j.ijfoodmicro.2015.04.017

Peptide nucleic acid fluorescence in-situ hybridization for identification of Vibrio spp. in aquatic products and environments

Int J Food Microbiol. 2015 Aug 3:206:39-44. doi: 10.1016/j.ijfoodmicro.2015.04.017. Epub 2015 Apr 17.

Authors

Xiaofeng Zhang¹, Ke Li², Shan Wu², Jiangbing Shuai², Weihuan Fang³

Affiliations

¹ 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.
² Food Safety Key Laboratory of Zhejiang Province, Zhejiang Entry-Exit Inspection and Quarantine Bureau, 126 Fuchun Road, Hangzhou 310012, People's Republic of China.
³ 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.

PMID: 25955286
DOI: 10.1016/j.ijfoodmicro.2015.04.017

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.

Keywords: Fluorescent in situ hybridization; Peptide nucleic acid; Vibrio species.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Typing Techniques / methods*
Bacterial Typing Techniques / standards
Environmental Microbiology*
Food Microbiology
In Situ Hybridization, Fluorescence*
Peptide Nucleic Acids / chemistry*
RNA, Ribosomal, 16S / genetics
Real-Time Polymerase Chain Reaction
Sensitivity and Specificity
Vibrio / genetics
Vibrio / physiology*
Water Microbiology*

Substances

Peptide Nucleic Acids
RNA, Ribosomal, 16S