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  

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

作　　者：向稚丹[1] 方峰[1] 舒赛男[1] 陈如[1] 聂兴草[1] 李革[1] 

机构地区：[1]华中科技大学同济医学院同济医院,武汉430030

出　　处：《中国实用儿科杂志》2005年第5期291-293,共3页Chinese Journal of Practical Pediatrics

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IS6110PCR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IS6110PCR检测系统可有效提高结核性脑膜炎诊断的阳性率。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.Results The 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.

关 键 词：结核性脑膜炎 脑脊液 二甲基亚砜 聚合酶链式反应 

分 类 号：R72[医药卫生—儿科]