Cooksey R, Morlock G, Abbadi S, Holloway B, Crawford J; American Society for Microbiology. General Meeting.
Abstr Gen Meet Am Soc Microbiol. 1997 May 4-8; 97: 546 (abstract no. U17).
National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA.
Among 367 Mtb isolates obtained during a one-month survey (April 1994) in New York City, 53 isolates (14.4%) were rifampin-resistant (RIFr). Nine distinct mutations in a 69-bp hypervariable region of rpoB were identified by automated sequence analyses among the RIFr isolates. PCR products (128-bp) encompassing this region were evaluated for electrophoretic mobility patterns using non-radioactive and fluorescent single-strand conformation polymorphism (SSCP analyses. Products evaluated by the former method were denatured in the presence of methyl mercury hydroxide and electrophoresed in precast acrylamide gels. Fluorogenically labeled primers were used to generate products for the latter method which were electrophoresed in an ABI Prism 310 Genetic Analyzer equipped with a 3% GeneScan Polymer Column. Mobility shifts for all nine mutations were clearly discernible from the wild type pattern by both methods. Fluorescent SSCP further enabled quantitation of mobility patterns and detection of populations containing both mutant and wild type products. These nonisotopic SSCP methods may serve as useful screens for RIF resistance in Mtb.
Publication Types:
Keywords:
- Mutation
- Mycobacterium tuberculosis
- New York City
- Polymerase Chain Reaction
- Polymorphism, Single-Stranded Conformational
- Rifampin
- genetics
Other ID:
UI: 102235433
From Meeting Abstracts