双色实时荧光PCR方法结合标本混合检测方案快速筛查肠道传染病菌(1)
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2010年11月5日
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[摘要] 目的：采用双色实时荧光PCR方法结合标本混合检测方案快速筛查检测公共场所从业人员肠道传染病菌，并与经典的培养法比较，探讨该方法的可行性。方法：随机采集5 000份从业人员的肛拭子标本，分别采用双色实时荧光PCR方法结合标本混合检测方案和“金标准”培养法同时检测肠道沙门和志贺菌，比较两种方法检测结果之间的差异。结果：双色实时荧光PCR方法结合标本混合检测方案的灵敏度为100%，特异性为99.64%，同培养法相比，具有省时、省力、操作安全、成本接近等优点，两种检测方法的阳性率无显著性差异。结论：提示双色实时荧光PCR方法结合标本混合检测方案可做为从业人员肠道传染病菌快速筛查的技术方法。
[关键词] 双色实时荧光PCR；标本混合检测方案；特异度；灵敏度；阳性率
[中图分类号] R446.5 [文献标识码] B[文章编号] 1674-4721(2010)11(a)-078-03
Rapid detection of intestinal pathogenes by real time PCR method combined with sample pooling process
LI Yiqiang, YANG Furong, KONG Minling, QIN Xiaojie, MAI Jiemei
(Center for Disease Control and Prevention of PanYu District in Guang Zhou City, Guangdong Province, Guang Zhou 511400, China)
[Abstract] Objective: The real time PCR method combined with sample pooling process was used to detect the intestinal pathogen in public sanitation related employees, its potential application was evaluated by compared with classical culture method. Methods: 5 000 anal swab specimens from public sanitation related employee were randomly collected, the samples were detected simultaneously for Salmonella and Shigella by real time PCR method combined with sample pooling process and "gold standard" culture method, the sensitivity and specificity of two were also evaluated. Results: The sensitivity and specificity of real time PCR method were 100% and 99.64% respectively, the positive rate showed no significant differences between two methods, however, the real time PCR method was faster, more secure and less laborious consuming compared with culture method. Conclusion: Real time PCR method combined with sample pooling process can be applied on the routine detection of intestinal pathogen in public sanitation related employee.
[Key words] Real time PCR; Sample pooling; Sensitivity; Specificity; Positive rate
目前检测从业人员肠道传染病菌的指标菌沙门氏菌和志贺氏菌的传统国家标准检验方法“金标准”培养法，其优点在于特异性强，但缺点是检测时间长，操作繁琐，灵敏度低，要求检验人员有丰富的工作经验。传统国家标准检验方法须5～7 d才能检出病原体，存在检验时间较长的问题。双色实时荧光PCR方法结合标本混合检测方案，是将双色实时荧光PCR方法技术[1]和标本混合检测方案技术[2]这两种现代先进技术结合在一起的一种新的技术方法。为探讨双色实时荧光PCR方法结合标本混合检测方案检测从业人员肠道传染病菌的可行性，随机采集5 000份从业人员的肛拭子标本，采用双色实时荧光PCR方法结合标本混合检测方案和“金标准”培养法同时检测肠道传染病菌，比较两种方法检测结果之间的差异，结果如下：
1 资料与方法
1.1 一般资料
随机抽取2009年6月～11月公共场所从业人员的5 000份肛拭子标本。
1.2 试剂
深圳生科源技术有限公司生产的沙门氏菌和志贺氏菌双色实时荧光PCR检测试剂、含营养肉汤增菌液的一次性多功能采样器 ......
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