DETECTION METHOD OF RIFAMPIN-RESISTANT MYCOBACTERIUM TUBERCULOSIS BY NESTED PCR-SSCP AND SINGLE STEP NESTED PCR-SSCP TARGETING RPOB GENE

摘要

PURPOSE: A detection method of Rifampin-resistant M. tuberculosis by nested PCR-SSCP and single step nested PCR-SSCP targeting a rpoB DNA fragment of the M. tuberculosis is provided which can prevent a false positive reaction by no tubercle acid-fast bacteria(MOTT), and can save time and expenses by treating many specimens. CONSTITUTION: The detection method of Rifampin-resistant M. tuberculosis is provided by outer PCR using an outer primer, which amplifies only rpoB DNA fragment(205bp) of the M. tuberculosis specifically without amplifying other bacterial rpoB DNA, nested PCR-SSCP(polymerase chain reaction-single strand conformational polymorphism) using an inner primer, which amplifies only rpoB DNA fragment(157bp) of the M. tuberculosis specifically, and single step nested PCR-SSCP. It takes about four days to detect the Rifampin-resistant M. tuberculosis by this method while it takes about twelve weeks by a conventional susceptibility test through a bacterial culture.