4重荧光PCR技术在霍乱监测与菌株鉴定的应用研究  

Application study of quadruplex real-time PCR surveillance and the strain identification for Vibrio cholerae

作　　者：黄仕杰[1] 李莉[1] 温慧欣[1] 黄建炜[1] 

机构地区：[1]厦门市疾病预防控制中心,福建361021

出　　处：《海峡预防医学杂志》2011年第5期1-2,65,共3页Strait Journal of Preventive Medicine

基　　金：卫生部-福建省医教联合攻关项目(No.WKJ2008-2-39);厦门卫生局科技计划项目(No.WSK05)

摘　　要：目的应用4重荧光PCR技术检测霍乱弧菌并对菌株进行鉴定。方法用PCR法对2008—2010年监测的水样、水产品标本及食物中毒疑似菌株以进行检测,同时用细菌学方法分离菌株,再进行PCR检测。结果 1 606份标本中,PCR法检出O1群霍乱弧菌阳性标本188份,从中分离到菌株150份(79.8%),O139群霍乱弧菌的PCR检测结果和分离结果均为阴性,其符合率为97.6%(1 568/1 606);用PCR法发现大量水产品带有非O1非O139群霍乱弧菌而ctxA阴性,但从中分离到1株ctxA阳性的菌株。结论 4重荧光PCR技术检测效果良好,灵敏度高于常规细菌学方法,可在霍乱弧菌检验中作为筛检和菌株鉴定的快速方法。ObjectiveTo detect Vibrio cholerae by quadruplex real-time PCR assay and identify the strains.MethodsThe V.choleraewere detected from 2008 to 2010 collected water,aquatic products and food-poisoning case samples by quadruplex real-time PCR assay and the stains were isolated and parallel study with classical bacteriological method to isolate the strains.And the isolated strains were screened and identified with quadruplex real-time PCR assay.ResultsTotally 188 in 1 606 samples were positive for O1 signal in PCR assay,but 150 strains（79.8%） of V.choleraeO1 were isolated with the classical method.V.choleraeO139 strains were tested negative neither with the PCR assay or the classical method.The coincidence rate between them was 97.6%（1 568/1 606）.And many sea food samples were tested positive for non-O1 and non-O139 and most of them were ctxA-negative,but one of them was ctxA-positive.ConclusionThe quadruplex real-time PCR is more significantly sensitive than classical one for V.choleraedetection in fast screening and further identifying V.cholerae in routine surveillance system.

关 键 词：霍乱弧菌 荧光PCR技术 快速筛检 菌株鉴定 

分 类 号：R378.3[医药卫生—病原生物学]