【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。更多还原

【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.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.更多还原