一种基于gyrB基因的用于检测创伤弧菌的聚合酶链式反应及其在牡蛎增菌液中直接检测该病原体的应用。
A gyrB-based PCR for the detection of Vibrio vulnificus and its application for direct detection of this pathogen in oyster enrichment broths.
作者信息
Kumar H Sanath, Parvathi Ammini, Karunasagar Indrani, Karunasagar Iddya
机构信息
Department of Fishery Microbiology, Karnataka Veterinary, Animal and Fisheries Sciences University, College of Fisheries, Mangalore 575 002, India.
出版信息
Int J Food Microbiol. 2006 Oct 1;111(3):216-20. doi: 10.1016/j.ijfoodmicro.2006.05.007. Epub 2006 Jul 18.
A polymerase chain reaction (PCR) method based on the gyrB (encoding gyrase B or topoisomerase II) gene sequence was developed for the detection of Vibrio vulnificus in seafood. The gyrB primers detected all laboratory isolates of V. vulnificus and did not cross react with other Vibiro and non-Vibrio species examined in this study. The sensitivity of detection of V. vulnificus by gyrB PCR was 300 CFU/g in artificially seeded oyster homogenate without enrichment while, 30 CFU/g could be detected following 18 h enrichment in alkaline peptone water (APW). The gyrB-specific PCR was employed for the direct detection of V. vulnificus in oyster enrichment broths. The assay detected V. vulnificus in 75% of natural oyster samples after 18 h enrichment in APW. The gyrB-based PCR described here offers a simple and specific one step PCR method for the detection of V. vulnificus in seafood enrichment broths.
开发了一种基于gyrB（编码促旋酶B或拓扑异构酶II）基因序列的聚合酶链反应（PCR）方法，用于检测海鲜中的创伤弧菌。gyrB引物可检测到所有创伤弧菌实验室分离株，且与本研究中检测的其他弧菌和非弧菌物种无交叉反应。在未经富集的人工接种牡蛎匀浆中，gyrB PCR检测创伤弧菌的灵敏度为300 CFU/g，而在碱性蛋白胨水（APW）中富集18小时后，可检测到30 CFU/g。gyrB特异性PCR用于直接检测牡蛎富集肉汤中的创伤弧菌。在APW中富集18小时后，该检测方法在75%的天然牡蛎样本中检测到了创伤弧菌。本文所述的基于gyrB的PCR为检测海鲜富集肉汤中的创伤弧菌提供了一种简单且特异的一步PCR方法。
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