副溶血性弧菌的免疫捕获PCR检测  

Detection of Vibio parahemolyticus using immunocaptured PCR

作　　者：郭川[1] 林洁敏 伦镜盛[2] 胡忠[2] 林耀盛[3] 方小衡[3] 

机构地区：[1]广东省汕头市疾病预防控制中心,广东汕头515041 [2]汕头大学生物学系,广东汕头515063 [3]广东药学院,广州510006

出　　处：《中国卫生检验杂志》2010年第6期1403-1405,共3页Chinese Journal of Health Laboratory Technology

基　　金：广东省科技计划项目(2006B20501008;2009B020309007);广东省医学科研基金项目(B2008161);广东省海洋渔业科技推广项目(A200901H06)

摘　　要：目的:将免疫捕获PCR法应用于副溶血性弧菌检测。方法:将脱毒后的副溶血性弧菌作为抗原免疫家兔,得到抗血清包被PCR管,对待检的副溶血弧菌进行免疫捕获,利用捕获到的菌体作为模板进行PCR检测。结果:PCR体系对副溶血性弧菌的tlh基因扩增具有特异性;抗体对副溶血弧菌起到一定程度的富集作用,包被抗血清后使PCR扩增出来的阳性结果更为明显,而抗血清本身没有扩增出DNA条带。结论:该检测方法能够对副溶血性弧菌进行特异性的检测,同时包被抗体的捕获对提高检测灵敏度起到一定的作用。Objective：To detect the Vibio parahemolyticus by using the immunocaptured PCR test.Methods：Vibio parahemolyticus was disactivated by formaldehyde and used as antigen to immune the rabbits.PCR tube was coated with the antiserum of the rabbits to capture the suspected bacteria.Afterwards,PCR was carried out to detect the captured bacteria.Results：The antibody in this antiserum shew the ability to enrich Vibio parahemolyticus,The PCR test which carry out in the serum coated tube and adding Vibio parahemolyticus cells as the template shew a positive result more significantly than the none coated tube,while the PCR test in the serum coated tube shew a negetive result without adding the template.Conclusion：The result reveals that the detection system can show high sepciallity in Vibio parahemolyticus test.Moreover,the antibody played a role in captureing the bacteria and promoting the sensitivity of the test.

关 键 词：副溶血性弧菌 免疫捕获PCR 抗血清 

分 类 号：R378.3[医药卫生—病原生物学]