Low-stringency single-specific-primer PCR as a tool for detection of mutations in the rpoB gene of rifampin-resistant Mycobacterium tuberculosis.
By the low-stringency single-specific-primer PCR technique, a highly sensitive and rapid method for diagnosis of rifampin resistance in Mycobacterium tuberculosis was established. Seven rifampin-resistant and five rifampin-susceptible specimens were analyzed. Rifampin resistance was determined by MIC measurement. A complex electrophoretic pattern consisting of many bands was obtained for both susceptible and rifampin-resistant isolates. The same pattern was obtained for all of the susceptible specimens, but differences between resistant and susceptible isolates were found. DNA sequencing showed that a particular mutation produces a specific electrophoretic pattern.
Carvalho
Wania S
WS
Departamento de Farmácia Social, Faculdade de Farmácia, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil. wscarv@icb.ufmg.br
Spindola de Miranda
Silvana
S
Costa
Kátia M
KM
Araújo
José G V C
JG
Augusto
Claudio J
CJ
Pesquero
João B
JB
Pesquero
Jorge L
JL
Gomes
Maria A
MA
D000904:Antibiotics, Antitubercular / Q000494:pharmacology*; D001483:Base Sequence; D017931:DNA Primers; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000145:classification / Q000187:drug effects* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012293:Rifampin / Q000494:pharmacology*; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D014397:Tuberculosis, Pulmonary / Q000382:microbiology
D000904:Antibiotics, Antitubercular; D017931:DNA Primers; D012321:DNA-Directed RNA Polymerases; D012293:Rifampin