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KMID : 0350519950480030919
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Journal of Catholic Medical College
1995 Volume.48 No. 3 p.919 ~ p.925
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Diagnosis of Tuberculosis by Polymerase Chain Reaction and Southern Blot
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Abstract
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Recently, mycobacterial infection has been increased sue partly to increase of human immunodeficiency virus infection. Therefore, rapid and precise diagnosis of tuberculosis is important. Although a presumptive diagnosis of tuberculosis can be
made
on
the basis of clinicl and radiological findings, isolation of the causative organism is required for the definitive diagnosis. However, routine culture of M. tuberculosis are very time-consuming. Recent advances in DNA techniques provided a rapid
diagnosis of tuberculosis by polymerase chain reaction (PCR) and/or Southern blot test.
To evaluate PCR and Southern blot for diagnosis of tuberculosis, part of pPH7301 was used for PCR and the 158 by PCR product was used as a probe for Southern blot test. The EcoR1, BstEⅡand PvuⅡwere used as restriction enzymes. The mycobacterial
species used in the study were 4 species of MOTT (Mycobacteria other than tuberculosis), and 9 strains of M. tuberculosis. Using PCR, M. tuberculosis and M. intracellulare revealed an amplified fragment of 158 bp. The fragment was absent in other
species. Using Southern blot test, only M. tuberculosis hybridized with 158 bp probe, and other species did not hybridized with the 158 bp probe. Southern blot analysis of EcoR1-digested DNA from 8 strains of M. tuberculosis revealed two
different
banding patterns, but different epidemiologic source was not found between these two groups. The similar banding patterns between these patients did not suggest an epidemiological reiationship, also. After restriction enzyme digestion, Southen
blot
test
exhibited different banding patterns.
Is conclusion, the PCR and Southern blot test are useful techique for rapid, sensitive, and definitive diagnosis of tuberculosis. Furthermore, this 158 bp fragment of pPH7301 could be useful for gtudy of tuberculosis.
Key words : polymerase chain reaction, Southern blot, M. tuberculosis
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KEYWORD
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