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| 副溶血弧菌和溶藻弧菌双重PCR检测方法的建立与初步应用 | |
| 引用本文： | 刘阳,孔繁德,徐淑菲,吴德峰,林立.副溶血弧菌和溶藻弧菌双重PCR检测方法的建立与初步应用[J].中国预防兽医学报,2012,34(8):647-650. |
| 作者姓名： | 刘阳 孔繁德 徐淑菲 吴德峰 林立 |
| 作者单位： | 1. 厦门出入境检验检疫局,福建厦门361026;福建农林大学动物科学学院,福建福州350002 2. 厦门出入境检验检疫局,福建厦门,361026 3. 福建农林大学动物科学学院,福建福州,350002 |
| 基金项目： | 国家质量监督检验检疫总局科技计划项目 |
| 摘 要： | 为建立同时快速检测海产品中的副溶血弧菌(VP)和溶藻弧菌(VA)的双重PCR方法,本研究根据VP和VA的toxR基因序列设计针对这两种细菌的两对特异性引物,建立能够快速同时检测这两种细菌的双重PCR方法,并对该反应体系的特异性和灵敏度进行检测。结果显示纯培养细菌VP和VA的检测灵敏度分别为2.32×103cfu/mL和2.56×103cfu/mL,临床病料检测灵敏度分别为2 cfu和3 cfu;与枸橼酸杆菌、沙门氏菌、美人鱼弧菌、霍乱弧菌、麦氏弧菌无交叉反应。研究表明本实验方法操作简便快速、特异性强、灵敏度高、稳定性好,并且经济实惠,值得推广应用。 |
| 关 键 词： | 副溶血弧菌 溶藻弧菌 双重PCR |
Establishment and primary application of duplex PCR for detection of Vibrio parahaemolytious and Vibrio alginolyticus |
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| LIU Yang , KONG Fan-de , XU Shu-fei , WU De-feng , LIN Li.Establishment and primary application of duplex PCR for detection of Vibrio parahaemolytious and Vibrio alginolyticus[J].Chinese Journal of Preventive Veterinary Medicine,2012,34(8):647-650. | |
| Authors: | LIU Yang KONG Fan-de XU Shu-fei WU De-feng LIN Li |
| Institution: | 1(1.Xiamen Entry-Exit Inspection and Quarantine Bureau,Xiamen 361026,China; 2.Animal Science College,Fujian Agricultural and Forestry University,Fuzhou 350002,China) |
| Abstract: | In this study,a duplex PCR(dPCR) was established and optimized to detect Vibrio parahaemolytious(VP) and Vibrio alginolyticus(VA) simultaneously,two sets of specific primers were designed according to the toxR sequence of VP and VA available in GenBank.The results showed that the detection limits of the dPCR assay were 2.32×103 cfu/mL and 2.56×103 cfu/mL for VP and VA with pure bacteria culture,and 2 cfu and 3 cfu for VP and VA with clinic samples,respectively.The method possessed a better specificity and no cross-reaction with other enterobacteria such as Citro Bacter,Salmonella,V.damsela,V.cholera,V.mets-chnikovii.The method could provide a useful assay for identification and inspection of VP or VA from fishery product in import and export. |
| Keywords: | Vibrio parahaemolytious Vibrio alginolyticus duplex polymerase chain reaction |
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