| 实时荧光PCR快速检测粪便中艰难梭菌方法 |
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| 引用本文： | 邵景东,吴琳,王毅谦,傅春玲,吴福平.实时荧光PCR快速检测粪便中艰难梭菌方法[J].中国卫生检验杂志,2011(7). |
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| 作者姓名： | 邵景东 吴琳 王毅谦 傅春玲 吴福平 |
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| 作者单位： | 张家港出入境检验检疫局;苏州大学医学部放射医学与公共卫生学院; |
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| 基金项目： | 江苏检验检疫局科研项目(2010KJ17) |
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| 摘 要： | 目的:建立实时荧光PCR快速检测艰难梭菌的方法。方法:以艰难梭菌磷酸丙糖异构酶(tpi)基因的保守序列为模板设计和合成特异性引物和荧光标记探针,建立实时荧光PCR检测体系,通过检测含有艰难梭菌标准菌株浓度为106-10 CFU/ml的细菌培养物及加标模拟样本进行敏感性分析,并对其特异性和干扰性进行评价。结果:该方法只对艰难梭菌进行特异性扩增,其他常见的病原菌均不能扩增;整个检测过程只需要2 h,对艰难梭菌菌悬液可检测至10 CFU/ml细菌,对加标粪便样本可检测至1000 CFU/ml细菌。结论:本研究建立的实时荧光PCR检测艰难梭菌方法具有快速、特异、敏感性高等优点,能实现对艰难梭菌的快速检测。
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| 关 键 词： | 艰难梭菌 Taqman探针 tpi 检测 |
Rapid detection of clostridium difficile in human stool by real-time fluorescence PCR |
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| SHAO Jing-dong,WU Lin,WANG Yi-qian,FU Chun-ling,WU Fu-ping.Rapid detection of clostridium difficile in human stool by real-time fluorescence PCR[J].Chinses Journal of Health Laboratory Technology,2011(7). |
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| Authors: | SHAO Jing-dong WU Lin WANG Yi-qian FU Chun-ling WU Fu-ping |
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| Institution: | SHAO Jing-dong1,WU Lin2,WANG Yi-qian1,FU Chun-ling2,WU Fu-ping1(1.Zhangjiagang Entry-Exit Inspection and Quarantine Bureau,Zhangjiagang 215600,China,2.School of Radiation Medicine & Public Health,Medical College of Soochow University,Suzhou 215123,China) |
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| Abstract: | Objective: To develop a real-time fluorescence PCR assay for rapid detection of Clostridium difficile.Methods: The special tpi gene of C.difficile were amplified through designing special primers and TaqMan probes within the conserved and specific regions for this gene.In this way,a rapid and stable method of real-time PCR assay for the detection of C.difficile standard bacterial concentration with 106-10 cfu/ml was established.The specificity and sensitivity of PCR were also analyzed.By adding standard cul... |
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| Keywords: | Clostridium difficile Taqman probe tpi Detection |
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