Multiplex polymerase chain reaction-based assay for the specific detection of toxin-producing Vibrio cholerae in fish and fishery products

被引：12
|
作者：
Jeyasekaran, Geevaretnam [1 ,2 ]
Raj, Kannan Thirumalai [1 ,2 ]
Shakila, Robinson Jeya [1 ,2 ]
Thangarani, Albin Jemila [1 ,2 ]
Sukumar, Durairaj [1 ,2 ]
机构：
[1] Tamil Nadu Vet & Anim Sci Univ, Coll Fisheries, Dept Fish Proc Technol, Tuticorin 628008, India
[2] Tamil Nadu Vet & Anim Sci Univ, Res Inst, Tuticorin 628008, India
来源：
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY | 2011年 / 90卷 / 03期
关键词：
Multiplex PCR; Vibrio; toxigenic strains of V. cholerae; rpoA gene; Toxin genes ctxA and RtxA genes; PCR ASSAY; RAPID DETECTION; IDENTIFICATION; GENE; O1; HEMOLYSIN; NON-O1; VIRULENCE; STRAINS; WATER;
D O I：
10.1007/s00253-011-3175-9
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
A multiplex polymerase chain reaction (MPCR)-based assay was developed for the simultaneous detection of Vibrios using the genus-specific RNA polymerase subunit A (rpoA) gene and specific detection of toxin-producing Vibrio cholerae strains using two sets of primer based on cholera toxin subunit A (ctxA) and repeat in toxin subunit A (RtxA)-producing genes. The MPCR method developed is applicable to both the simultaneous and the two-step detection of genus Vibrio total and toxigenic V. cholerae species. This assay was specific as no amplification occurred with the other bacterial pathogens tested. The sensitivity of the assay was tested by artificially spiking the shrimp homogenate with the toxigenic strain of V. cholerae (NICED 16582) in different dilutions. The developed MPCR assay could detect three cells of V. cholerae in 12 h pre-enrichment in APW. The proposed method is rapid, sensitive, and specific for the detection of Vibrio genus as well as toxin-producing V. cholerae strains in environmental samples.
引用
收藏
页码：1111 / 1118
页数：8
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