Kocagöz T, Yilmaz E, Ozkara S, Kocagöz S, Hayran M, Sachedeva M, Chambers H F
Department of Microbiology, School of Medicine, Hacettepe University, Ankara, Turkey.
J Clin Microbiol. 1993 Jun;31(6):1435-8. doi: 10.1128/jcm.31.6.1435-1438.1993.
A repetitive sequence of Mycobacterium tuberculosis DNA was amplified by polymerase chain reaction (PCR), from sputum samples, for the diagnosis of pulmonary tuberculosis. The method of heating the sample in a boiling water bath to break down the bacterial cell wall and to release the DNA was compared with that of enzymatic lysis of bacteria and then phenol-chloroform extraction of DNA. Heating the sample was the better method with a sensitivity of approximately 10 microorganisms. A total of 78 sputum specimens prepared by heating were examined by PCR, and the results were compared with the results of acid-fast stained smears, cultures, and clinical data. M. tuberculosis was detected by PCR in all smear- and culture-positive and smear-negative, culture-positive cases. Additionally, PCR was capable of detecting four of nine cases which were smear and culture negative but clinically suspected of tuberculosis. DNA amplification by PCR is a sensitive and specific method for the diagnosis of tuberculosis, and with this simplified DNA isolation procedure it can be used in routine clinical practice.
采用聚合酶链反应（PCR）扩增结核分枝杆菌DNA的重复序列，用于从痰标本中诊断肺结核。将在沸水浴中加热标本以破坏细菌细胞壁并释放DNA的方法与细菌酶解后用酚-氯仿提取DNA的方法进行了比较。加热标本是更好的方法，灵敏度约为10个微生物。对78份经加热制备的痰标本进行了PCR检测，并将结果与抗酸染色涂片、培养结果及临床资料进行了比较。在所有涂片和培养均阳性以及涂片阴性、培养阳性的病例中，PCR均检测到结核分枝杆菌。此外，PCR能够检测出9例涂片和培养均阴性但临床怀疑为结核病的病例中的4例。通过PCR进行DNA扩增是诊断结核病的一种灵敏且特异的方法，采用这种简化的DNA分离程序，它可用于常规临床实践。
