Detection of Mycobacterium tuberculosis in Urine by Nested Polymerase Chain Reaction / 대한비뇨기과학회지

ABSTRACT

Today, genitourinary tuberculosis as well as tuberculosis of other organs have decreased in its incidence with development of antituberculosis drugs. However, it is still very frequent in Korea. There are still great deal of difficulties in finding Mycobacterium tuberculosis from urine because of variations in method of taking urine sample and techniques of examination. For these matters, early diagnosis was difficult indeed. For this reason, we performed amplification of Mycobacterial DNA from urine by use of the Polymerase Chain Reaction, and urine AFB smear from Apr., 1993 to Feb., 1994 and achieved following results. l. In order to increase sensitivity and specificity, we performed nest PCR. and primers used each cases were a) INS1 and INS2, which amplify 245-base pair sized DNA, b) Pt3 and Pt6, which amplify 188-base pair. The sequences of primers were INS1 (5'-CGTGAGGGCATCGAGGTGGC-3'), INS2 (5'- GCGTAGGCGTCGGTGACAAA-3'), Pt3(5'-GAACGGCTGATGACCAAACT-3'), and Pt6( 5'-ACGTAGGCGAACCCTGCCCA-3') (Table l). And after 30 cycles of amplification, unique Mycobacterium tuberculosis DNA band was found ( Figure). 2. Total number of confirmed genitourinary tuberculosis were 35 cases, 19 cases were positive in both PCR and AFB smear, other 15 cases were positive only in PCR and in one case, PCR was negative but positive in AFB smear (Table 2).