Mutations in the rpoB gene of Mycobacterium tuberculosis that interfere with PCR-single-strand conformation polymorphism analysis for rifampin susceptibility testing.
Rifampin susceptibility of 32 rifampin-resistant and 26 rifampin-susceptible Mycobacterium tuberculosis strains was analyzed by PCR-single-strand conformation polymorphism (SSCP) and DNA sequencing within the 157-bp region of the rpoB gene (Ala500 to Val550). Two false-positive PCR-SSCP results were observed among the susceptible strains due to the silent mutation Gln513 (CAA-->CAG) and the deletion mutation Thr508 and Ser509. Another silent mutation [Leu511 (CTG-->CTA)], combined with the mutation Ser531-->Leu, was observed in a resistant strain. These results suggest that to rule out false-positive PCR-SSCP results, sequencing of the target DNA is required.
Kim
B J
BJ
Department of Microbiology and Cancer Research Center, Seoul National University College of Medicine, Korea.
Kim
S Y
SY
Park
B H
BH
Lyu
M A
MA
Park
I K
IK
Bai
G H
GH
Kim
S J
SJ
Cha
C Y
CY
Kook
Y H
YH
D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D004352:Drug Resistance, Microbial; D005189:False Positive Reactions; D008826:Microbial Sensitivity Tests; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics*; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D012293:Rifampin / Q000494:pharmacology*; D017384:Sequence Deletion
D012321:DNA-Directed RNA Polymerases; D012293:Rifampin