中国实用儿科杂志

中国实用儿科杂志

• 论著 • 上一篇    下一篇

改良脑脊液处理方法和优化PCR检测系统对结核性脑膜炎患儿的诊断价值

向稚丹;方峰;舒赛男;陈如;聂兴草;李革   

  1. 华中科技大学同济医学院同济医院(武汉，430030)
  • 收稿日期:2004-11-12 修回日期:2005-02-27 出版日期:2005-05-25 发布日期:2005-05-25
  • 通讯作者: 向稚丹

Value of modified method of cerebrospinal fluid preparation and optimized PCR system in diagnosing tuberculous meningitis.

Xiang Zhidan,Fang Feng,Shu Sainan，et al.   

  1. Tongji Hospital,Tongji Medical College,Hua Zhong University of Science and Technology,Wuhan 430030,China
  • Received:2004-11-12 Revised:2005-02-27 Online:2005-05-25 Published:2005-05-25
  • Contact: Xiang Zhidan

摘要： 目的 改良脑脊液样本处理方法和优化PCR检测系统，提高结核性脑膜炎诊断阳性率。方法 建立微波-碱性非离子型表面活性剂-氯仿法，并用二甲基亚砜(DMSO)优化结核杆菌插入序列IS 6110 PCR检测系统；分别用微波-碱性非离子型表面活性剂-氯仿法和简易氯仿法处理1998年7月至2004年11月间华中科技大学同济医学院同济医院收集的108份脑脊液样本，经5%DMSO优化的PCR诊断系统扩增，将结果进行比较。结果 微波-碱性非离子型表面活性剂-氯仿法处理脑脊液模板PCR诊断结核性脑膜炎阳性率为88%，简易氯仿法为71%,假阳性率均为0。结论 微波-碱性非离子型表面活性剂氯仿法和5%DMSO优化的IS 6110 PCR检测系统可有效提高结核性脑膜炎诊断的阳性率。


Abstract Objective To improve the diagnosing level of tuberculosis meningitis (TBM) by means of modified preparation of cerebrospinal fluid (CSF) and optimized IS6110 PCR system.Methods Establishing the modified templatepreparing method of microwave/ alkaline nonionic detergent/ chloroform and using dimethyl sulphoxide (DMSO) to optimize the PCR system of tubercle bacillus(TB) gene IS6110.Preparing the template from 108 CSF specimens with the microwave/alkaline nonionic detergent/ chloroform method and amplifying IS 6110 gene with DMSO modified system,comparing with the simple chloroform method.ResultsThe modified IS 6110 PCR assay with microwave/alkaline nonionic detergent/chloroform method showed the positive rate of 88%,while the rate of simple chloroform method was 71%.Both false positive rates were 0%. Conclusion The method of preparing the template with the microwave/alkaline nonionic detergent/chloroform method and modifying TB IS 6110 PCR system with 5%DMSO is an effective way to increase the positive rate of TBM diagnosis.
Key words Tuberculosis meningitis;Cerebrospinal fluid;Dimethylul sphoxide;Polymerase chain reaction

关键词: 结核性脑膜炎, 脑脊液, 二甲基亚砜, 聚合酶链式反应

Abstract: Abstract Objective To improve the diagnosing level of tuberculosis meningitis (TBM) by means of modified preparation of cerebrospinal fluid (CSF) and optimized IS6110 PCR system.Methods Establishing the modified templatepreparing method of microwave/ alkaline nonionic detergent/ chloroform and using dimethyl sulphoxide (DMSO) to optimize the PCR system of tubercle bacillus(TB) gene IS6110.Preparing the template from 108 CSF specimens with the microwave/alkaline nonionic detergent/ chloroform method and amplifying IS 6110 gene with DMSO modified system,comparing with the simple chloroform method.ResultsThe modified IS 6110 PCR assay with microwave/alkaline nonionic detergent/chloroform method showed the positive rate of 88%,while the rate of simple chloroform method was 71%.Both false positive rates were 0%. Conclusion The method of preparing the template with the microwave/alkaline nonionic detergent/chloroform method and modifying TB IS 6110 PCR system with 5%DMSO is an effective way to increase the positive rate of TBM diagnosis.

Key words: Polymerase chain reaction , Cerebrospinal fluid, Dimethylul sphoxide