| 闫志勇,王斌,毕春霞.多重半套式聚合酶链反应在检测脑脊液病原菌中的应用[J].Chinese journal of Epidemiology,2003,24(4):296-299 |
| 多重半套式聚合酶链反应在检测脑脊液病原菌中的应用 |
| Polymerase chain reaction of multiple half sleeve type application in the detection of cerebrospinal fluid pathogen |
| Received:June 20, 2002 |
| DOI： |
| KeyWord: 聚合酶链反应 脑脊液 细菌 16SrRNA基因 |
| English Key Word: Polymerase chain reaction Cerebrospinal fluid bacteria 16 s rRNA genes |
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| Abstract: |
| 目的 建立多重半套式聚合酶链反应(PCR)快速检测脑脊液标本中常见的病原菌。方法 通过对病原菌 1 6SrRNA基因保守区和变异区的序列分析,设计通用引物及革兰阴性菌、革兰阳性菌的特异性引物,分别作为外、内侧引物,对脑脊液标本中不同细菌的DNA进行多重半套式PCR扩增 ;同时与常规细菌培养法作比较,并检测了该方法的敏感性。结果 外侧扩增后革兰阳性菌、革兰阴性菌经扩增后均有长约 1 0 32bp的片段产生 ;内侧扩增两种细菌除 1 0 32bp的产物外,革兰阳性菌另有一 336bp的特异性产物,革兰阴性菌另有一 1 2 7bp的特异性产物。该方法最低可检测出8cfu ml的大肠埃希菌 ;62份脑脊液标本扩增结果与培养法相比,敏感性、特异性、阳性预测值、阴性预测值分别为 93 .8%、5 .7%、88.2 %、97.8%。b>结论 多重半套式PCR方法能特异、敏感、快速地检测出脑脊液感染的常见病原菌 |
| English Abstract: |
| Purpose Purpose to establish multiple polymerase chain reaction (PCR) of half sleeve type rapid detection of common pathogenic bacteria in the cerebrospinal fluid specimens. Methods of pathogen 16 s rRNA genes conservative area and variation area sequence analysis, general design primers and gram-negative bacteria, gram-positive bacteria of specific primers, respectively as the outside and the inside of the primer, in cerebrospinal fluid specimens of different bacterial DNA for PCR amplification of multiple half sleeve type;At the same time compare to conventional bacterial culture method, and test the sensitivity of the method. Results The results of the lateral expansion after the gram-positive bacteria and gram-negative bacteria after amplification has about 1, 032 bp long pieces;Medial expansion two kinds of bacteria in addition to the product of 1 032 bp, gram-positive bacterium another 336 bp product specificity, the specificity of the gram-negative bacteria and 127 a bp. This method can detect the lowest 8 cfu/ml of e. coli;62 cerebrospinal fluid specimens amplification results compared with culture method, the sensitivity, specificity, positive predictive value, negative predictive value were 93.8%, 5.7%, 88.2%, 97.8%. Conclusion multiple half sleeve polymerase chain reaction (PCR) can detect specific, sensitive and quick common pathogenic bacteria of cerebrospinal fluid infections. |
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