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


KOCAGOZ T. , YILMAZ E., OZKARA S., KOCAGOZ S. , HAYRAN M., SACHEDEVA M., et al.

Journal of Clinical Microbiology, cilt.31, ss.1435-1438, 1993 (SCI İndekslerine Giren Dergi) identifier identifier identifier

  • Cilt numarası: 31 Konu: 6
  • Basım Tarihi: 1993
  • Dergi Adı: Journal of Clinical Microbiology
  • Sayfa Sayısı: ss.1435-1438

Özet

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.