Detection and identification of Vibrio parahaemolyticus by multiplex PCR and DNA-DNA hybridization on a microarray

In this paper, we developed a rapid and accurate method for the detection of Vibrio parahaemolyticus strains, using multiplex PCR and DNA--DNA hybridization. Multiplex PCR was used to simultaneously amplify three diagnostic genes （tlh, tdh andfla） that serve as molecular markers of V. parahaemolytic...

Full description

Saved in:
Bibliographic Details
Published in:Journal of genetics and genomics 2011-03, Vol.38 (3), p.129-135
Main Authors: Wang, Rongzhi, Huang, Jiadong, Zhang, Wei, Lin, Guangmei, Lian, Junwei, Jiang, Libin, Lin, Hongcong, Wang, Songfa, Wang, Shihua
Format: Article
Language:English
Subjects:
Alkaline phosphatase
DNA microarrays
DNA Probes
DNA杂交
Foodborne Diseases - microbiology
Gene microarray
Hemolysin Proteins - genetics
Humans
Hybridization
Multiplex PCR
Oligonucleotide Array Sequence Analysis - methods
PCR方法
Polymerase chain reaction
Polymerase Chain Reaction - methods
Primers
Vibrio
Vibrio Infections - diagnosis
Vibrio parahaemolyticus
Vibrio parahaemolyticus - genetics
Vibrio parahaemolyticus - isolation & purification
分子标记
副溶血性弧菌
多重PCR
微阵列
检测
鉴定
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:In this paper, we developed a rapid and accurate method for the detection of Vibrio parahaemolyticus strains, using multiplex PCR and DNA--DNA hybridization. Multiplex PCR was used to simultaneously amplify three diagnostic genes （tlh, tdh andfla） that serve as molecular markers of V. parahaemolyticus. Biotinylated PCR products were hybridized to primers immobilized on a microarray, and detected by chemiluminesce with avidin-conjugated alkaline phosphatase. With this method, forty-five samples were tested. Eight known virulent strains （tlh＋/tdh＋/fla＋） and four known avirulent strains （tlh＋/tdh /fla＋） of the V. parahaemolyticus were successfully detected, and no non-specific hybridization and cross-hybridization reaction were found from fifteen closely-related strains （tlh-/tdh-/fla＋） of the Vibrio spp. In addition, all the other eighteen strains of non-Vibrio bacteria （tlh-/tdh /fla-） gave negative results. The DNA microarray successfully distinguished V. parahaemolyticus from other Vibrio spp. The results demonstrated that this was an efficient and robust method for identifying virulent strains of V. parahaemolyticus.
ISSN:1673-8527
DOI:10.1016/j.jgg.2011.02.002