PCR-Restriction Fragment Length Polymorphism Analysis for Identification of Bacteroides spp. and Characterization of Nitroimidazole Resistance Genes
AUTOR(ES)
Stubbs, Simon L. J.
FONTE
American Society for Microbiology
RESUMO
Bacteroides spp. are opportunist pathogens that cause blood and soft tissue infections and are often resistant to antimicrobial agents. We have developed a combined PCR-restriction fragment length polymorphism (RFLP) technique to characterize the 16S rRNA gene for identification purposes and the nitroimidazole resistance (nim) gene for detection of resistance to the major antimicrobial agent used to treat Bacteroides infections: metronidazole (MTZ). PCR-RFLP analysis of 16S ribosomal (rDNA) with HpaII and TaqI produced profiles that enabled discrimination of type strains and identification of 70 test strains to the species level. The 16S rDNA PCR-RFLP identification results agreed with routine phenotypic testing for 62 of the strains. The discrepancies between phenotypic and PCR-RFLP methods for eight strains were resolved by 16S rDNA sequencing in three cases, but five strains remain unidentified. The presence of nim genes was indicated by PCR in 25 of 28 strains that exhibited reduced sensitivity to MTZ. PCR-RFLP of the nim gene products identified the four reported genes (nimA, -B, -C, and -D) and indicated the presence of a previously unreported nim gene in 5 strains. This novel nim gene exhibited 75% DNA sequence similarity with nimB. These rapid, accurate, and inexpensive methods should enable improved identification of Bacteroides spp. and the detection of MTZ resistance determinants.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=87358Documentos Relacionados
- Identification of Staphylococcus spp. by PCR-Restriction Fragment Length Polymorphism of gap Gene
- PCR-Restriction Fragment Length Polymorphism Analysis for Detection of Point Mutations Associated with Macrolide Resistance in Campylobacter spp.
- Identification of Oral Peptostreptococcus Isolates by PCR-Restriction Fragment Length Polymorphism Analysis of 16S rRNA Genes
- Routine use of PCR-restriction fragment length polymorphism analysis for identification of mycobacteria growing in liquid media.
- Development of a Species-Specific PCR-Restriction Fragment Length Polymorphism Analysis Procedure for Identification of Madurella mycetomatis