[Application of PCR-SSCP technique in detection of rpoB gene mutation in rifampin-resistant Mycobacterium tuberculosis].
OBJECTIVE
To evaluate rpoB gene mutation in rifampin-resistant Mycobacterium tuberculosis and its relationship with rifampin resistance.


METHODS
Forty clinical isolates of Mycobacterium tuberculosis were analyzed by PCR-SSCP technique, with H37Rv reference strains as control group.


RESULTS
The sensitivity of amplication products of 411bp and 258bp were found to be 5 pg/microliters, 500 organisms per milliliter and 1 pg/microliter, 500 organisms per milliliter respectively. rpoB gene belongs to genus specificity. Characteristics of SSCP graph of 258bp fragment: Ten sensitive strains were the same as H37 Rv. Thirty strains of rifampin-resistant or multidrug resistance, including rifampin, were different from H37Rv except for three strains. Positive rate was 90%, while specificity 100%.


CONCLUSIONS
Results showed that PCR-SSCP technique could detect rPOB gene mutation, which might associate with rifampin resistance and be helpful to rapid detection and research of rifampin-resistant Mycobacterium tuberculosis.
He
X
X
Tuberculosis Research Laboratory, 309th Hospital of PLA, Beijing.
Zhuang
Y
Y
Li
G
G
D000904:Antibiotics, Antitubercular / Q000494:pharmacology; D004352:Drug Resistance, Microbial / Q000235:genetics; D005798:Genes, Bacterial; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000302:isolation & purification; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D012319:RNA Polymerase II / Q000235:genetics*; D012293:Rifampin / Q000494:pharmacology; D014397:Tuberculosis, Pulmonary / Q000382:microbiology
D000904:Antibiotics, Antitubercular; D012319:RNA Polymerase II; D012293:Rifampin