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. 1993 Jun;31(6):1435–1438. doi: 10.1128/jcm.31.6.1435-1438.1993

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

T Kocagöz 1, E Yilmaz 1, S Ozkara 1, S Kocagöz 1, M Hayran 1, M Sachedeva 1, H F Chambers 1
PMCID: PMC265557  PMID: 8314982

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.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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