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J Clin Microbiol. 1993 June; 31(6): 1435-1438

Detection of Mycobacterium tuberculosis in sputum samples by polymerase chain reaction using a simplified procedure.

T Kocagöz, E Yilmaz, S Ozkara, S Kocagöz, M Hayran, M Sachedeva and H F Chambers

Department of Microbiology, School of Medicine, Hacettepe University, Ankara, Turkey.

ABSTRACT

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.

J Clin Microbiol. 1993 June; 31(6): 1435-1438




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