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KMID : 0357319930280020131
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Journal of the Korean Society for Microbiology
1993 Volume.28 No. 2 p.131 ~ p.142
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Detection of Mycobacterium tuberculosis in Sputum Samples by Polymerase Chain Reaction
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조은경
최대경/백태현/박정규/김화중
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Abstract
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The polymerase chain reaction (PCR) was used to detect M. tuberculosis DNA in sputum samples from 126 tuberculosis. The target DNA is a 123-base pair(bp) segment of IS6110, which is repeated in the M. tubercuylosis chromosome and specific for the
M.
tuberculosis complex. Amplified products were detected by gel electophoresis and by Southern blot hybridization with 32P-labeled 123-bp DNA probe.
Fifty-nine of 126 patients were active tuberculosis in which were confirmed M. buberculosis by culture and staining method. All of these active tuberculosis patients showed the PCR positive products. 33 of 66 inactive tuberculosis patients who
have
bacteriologically negative results showed the PCR positibe products. Among 33 PCR negative specimens, 30 cases were inactive stage being treated with anti-tuberculosis drugs for six months or loger, 3 cases treated for six months and their
medication
were ceased. 14 cases of total specimens were obtaied from patients initialy suspected tuberculosis because of the presence of clinical symptoms or compatible abnormalities on chest radiograph and these were all PCR positive.
The cell lysis huffer treatment method was more sensitive than Triton X-100 treatment method. By the cell lysis buffer method, one to ten bacteria were dctectable by ethidium bromide staining. And the detecton of PCR producs by Southern blot
hybridization was 12*-10* fold more sensitive than that by ethidium bromide staining. 12 of 25 PCR negative samples by ethidium-bromide staining were ultimately confirmed PCR positive by Southern blot hybridization.
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