微波-碱性非离子型表面活性剂-氯仿法用于结核性脑膜炎脑脊液模板处理的效果观察

Effect of the microwave/ alkaline nonionic detergent/ chloroform method on cerebrospinal fluid preparation for diagnosing tuberculosis meningitis by PCR.

目的观察微波-碱性非离子型表面活性剂-氯仿法用于结核性脑膜炎脑脊液模板处理的效果。方法建立微波-碱性非离子型表面活性剂-氯仿法用于结核性脑膜炎脑脊液模板处理,并与简易氯仿法、碱性非离子型表面活性剂法和经典酚氯仿法比较;用PCR抑制物实验确定脑脊液中抑制PCR反应的成分。结果同一份结核杆菌培养阳性的脑脊液样本微波-碱性非离子型表面活性剂-氯仿法检出的最低稀释度为1∶8,简易氯仿法为1∶4,碱性非离子型表面活性剂法和经典酚氯仿法均为1∶2;结核杆菌阳性脑脊液上清液抑制PCR反应,氯仿抽提可去除抑制物。结论微波-碱性非离子型表面活性剂-氯仿法处理模板可有效提高结核杆菌IS6110PCR扩增的敏感性。

Objective To improve the detection rate of tuberculosis meningitis（TBM） with modified preparation of cerebrospinal fluid（CSF）. Methods ①Establishing the microwave/alkaline nonionic detergent/chloroform method for CSF specimens＇ preparation, and comparing with the simple chloroform method, the alkaline nonionie detergent method and the phenol/ chloroform precipitation method. ②Utilizing the amplification system of a house-keeping gene to identify the PCR inhibitory factor in the CSF of TBM. Results ①Using the same CSF sample, whose TB was positive proved by euhure, the microwave/ alkaline nonionie detergent/chloroform method for IS6110 PCR system showed the lowest detectable dilution, 1：8. The simple chloroform method was 1 ： 4. The alkaline-nonionie detergent method and the phenol/chloroform precipitation method were both 1：2. ②Extracting with chloroform could remove the PCR inhibitory factor, which were contained in the CSF of TBM. Conclusion Preparing the template with the microwave/alkaline nonionie detergent/chloroform method is effective for improving the positive rate of TBM diagnosis.