Rapid detection of Vibrio parahaemolyticus in seafood by Real- time PCR targeted to the H- NS gene

ZHU Yao-lei; SANG Xue; GAN Wei-qi; ZHANG Gong-liang; HOU Hong-man

doi:10.13386/j.issn1002-0306.2015.19.024

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Science and Technology of Food Industry > 2015 > (19): 155-158. > DOI: 10.13386/j.issn1002-0306.2015.19.024

ZHU Yao-lei, SANG Xue, GAN Wei-qi, ZHANG Gong-liang, HOU Hong-man. Rapid detection of Vibrio parahaemolyticus in seafood by Real- time PCR targeted to the H- NS gene[J]. Science and Technology of Food Industry, 2015, (19): 155-158. DOI: 10.13386/j.issn1002-0306.2015.19.024

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Rapid detection of Vibrio parahaemolyticus in seafood by Real- time PCR targeted to the H- NS gene

School of Food Science and Technology,Dalian polytechnic University

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Received Date: January 11, 2015

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Objective: In this study,a rapid and accurate method was established to detect Vibrio parahaemolyticus of using SYBR Green I fluorescence PCR reaction.Methods: Primers were designed by using conserved region of the H- NS gene.Twelve non Vibrio parahaemolyticus were amplified by PCR to confirm the specificity.Based on H- NS gene,the standard curve was established and the sensitivity was determined by SYBR Green I fluorescence PCR reaction. Sterile clam samples contaminated by different concentrations of bacteria were detected to determine the feasibility of this method.Then,thirty clam samples were detected,and the results were compared with the GB 4789.7-2013.Results: Correlation coefficient of the standard curve established in this study was 0.998,and the sensitivity was 7.3 × 10 CFU / m L.The detection limit of artificial contamination was 6.7 × 102 CFU / m L.There were four positive results detected in the thirty samples by using SYBR Green I fluorescence PCR method which was consistent with GB 4789.7- 2013.Conclusion: Method of this article could be used in rapid detection of Vibrio parahaemolyticus.
- Vibrio parahaemolyticus,
- SYBR Green I,
- Real-time PCR,
- H-NS gene

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[1]	李杰,卢昕,徐嘉良,等.应用SYBR Green I荧光PCR方法快速检测溶藻弧菌[J].中国预防医学杂志,2014,15(3):195-198.
[2]	覃倚莹,吴晖,肖性龙,等.toxR基因作为荧光定量PCR靶基因设计Taq Man探针快速检测副溶血弧菌[J].生物工程学报,2008,24(10):1837-1842.
[3]	徐晓可,吴清平,张菊梅,等.水产品中副溶血性弧菌特异性二重PCR检测方法的研究[J].食品与机械,2008,24(3):90-93.
[4]	Muhammad Tofazzal Hossain,Young-Ok Kim,In-Soo Kong.Multiplex PCR for the detection and differentiation of Vibrio parahaemolyticus strains using the gro EL,tdh and trh genes[J].Molecular and Cellular Probes,27(2013):171-175.
[5]	Tove Atlung,Hanne Ingmer.H-NS:a modulator of environmentally regulated gene expression[J].Molecular Microbiology,1997,24(1):7-17.A
[6]	R.No,K.Okada,K.Kogure,et al.Rapid detection of Vibrio parahaemolyticus by PCR targeted to the histone-like nucleoid structure(H-NS)gene and its genetic characterization[J].Letters in Applied Microbiology,2011,53:127-133.
[7]	王昊宇,张公亮,侯红漫.应用SYBR Green I溶解曲线检测食源性单增李斯特菌[J].食品工业科技,2013,34(10):77-79.
[8]	Anuj Tyagi,Saravanan V,Iddya Karunasagar,et al.Detection of Vibrio parahaemolyticus in tropical shellfish by SYBR green I real-time PCR and evaluation of three enrichment media[J].International Journal of Food Microbiology,2009,129(2):124-130.
[9]	Zhou S,Hou Z,Li N,et al.Development of a SYBR Green I real-time PCR for quantitative detection of Vibrio alginolyticus in seawater and seafood[J].Journal of Applied Microbiology,2007,103(5):1897-1906.
[10]	Amy V Rizvi,Asim K Bej.Multiplexed real-time PCR amplification of tlh,tdh and trh genes in Vibrio parahaemolyticus and its rapid detection in shellfish and Gulf of Mexico water[J].Antonie van Leeuwenhoek,2010,98:279-290.
[11]	Shuijing Yu,Wanyi Chen,Da peng Wang,et al.Speciesspecific PCR detection of the food-borne pathogen Vibrio parahaemolyticus using the irg B gene identified by comparative genomic analysis[J].FEMS Microbiol Letters,2010,30(7):65-71.
[12]	Yoo Seok Jeong,Hee Kyoung Jung,Joo-Heon Hong.Multiplex real-time polymerase chain reaction for rapid detection of Staphylococcus aureus,Vibrio parahaemolyticus,and Salmonella typhimurium in milk and kimbap[J].Journal of the Korean Society for Applied Biological Chemistry,2013,56(6):715-721.
[13]	SN/T 3924-2014,出口贝类中大肠菌群、粪大肠菌群检测方法[S].
[14]	SN/T 1194-2014,植物及其产品转基因成分检测[S].
[15]	Blackstone G M,Noalsmxn J L,Viekery M C,et al.Detection of pathogenic Vibrio parahaemolyticus in oyster enrichments by real-time PCR[J].Microbiol Methods,2003,53(2):149-155.
[16]	蔡潭溪,蒋鲁岩,黄克,等.基于Taq Man探针的Real—time PCR技术定量检测副溶血弧菌[J].微生物学报,2005,45(4):638-642.

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Rapid detection of Vibrio parahaemolyticus in seafood by Real- time PCR targeted to the H- NS gene

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