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结核性胸腔积液的实验诊断,目前尚无满意方法,临床诊断多为经验性的。聚合酶链反应(PCR)能对病原微生物的特异 DNA 序列作高效的体外扩增,是一类敏感、特异、快速的检测方法。胸水结核杆菌的 PCR 测定,国外仅有少数实例报告,我们采用国产试剂、仪器初步建立了这一方法。肝素抗凝胸水经低速离心除去部分混杂细胞,蒸馏水溶解红细胞,高速离心浓集结核杆菌,冻融3轮,使菌体裂解、释放 DNA,用蛋白酶 K 消化2小时,95℃加热10分钟灭活酶,高速离心,上清液即为 DNA 粗提液。
There are no satisfactory methods for the diagnosis of tuberculous pleural effusion. Clinical diagnosis is mostly empirical. Polymerase chain reaction (PCR) can efficiently amplify specific DNA sequences of pathogenic microorganisms and is a sensitive, specific and rapid detection method. PCR detection of Mycobacterium tuberculosis, only a few cases of foreign reports, we use domestic reagents, equipment initially established this method. Heparin anticoagulated pleural effusion by low speed centrifugation to remove some of the mixed cells, red blood cells were dissolved in distilled water, concentrated Mycobacterium tuberculosis by high-speed centrifugation, freeze-thaw 3, the cell lysate, DNA release, digestion with proteinase K 2 hours, 95 ℃ heating 10 minutes inactivation Enzyme, high-speed centrifugation, the supernatant is DNA crude extract.