RESUMO
A multiplex quantitative reverse transcription-PCR assay was developed to detect azole resistance in Candida glabrata, an important opportunistic pathogen that develops resistance rapidly. Resistance was defined as a >or=3-fold increase in CDR1 expression by this assay, which proved to be 100% sensitive and 95% specific in comparison to the gold standard broth microdilution assay.
Assuntos
Antifúngicos/farmacologia , Azóis/farmacologia , Candida glabrata/efeitos dos fármacos , Farmacorresistência Fúngica , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Vagina/microbiologia , Candida glabrata/isolamento & purificação , Candidíase Vulvovaginal/microbiologia , Feminino , Fluconazol , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Humanos , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Testes de Sensibilidade Microbiana/métodos , Sensibilidade e Especificidade , Regulação para CimaRESUMO
A retrospective survey of 93,775 samples testing positive in Candida species-specific PCR tests performed on cervicovaginal swabs over a 4-year period demonstrated consistent yearly distributions of Candida albicans (89%), C. glabrata (7.9%), C. parapsilosis (1.7%), and C. tropicalis (1.4%). However, the species distributions among different age groups revealed increases in the percentages of non-albicans species with increases in age.
Assuntos
Candida/classificação , Candida/isolamento & purificação , Candidíase Vulvovaginal/epidemiologia , Candidíase Vulvovaginal/microbiologia , Vagina/microbiologia , Adulto , Fatores Etários , Idoso , Candida/genética , Candida albicans/classificação , Candida albicans/genética , Candida albicans/isolamento & purificação , Colo do Útero/microbiologia , DNA Fúngico , Feminino , Humanos , Pessoa de Meia-Idade , Técnicas de Tipagem Micológica , Reação em Cadeia da Polimerase/métodos , Especificidade da Espécie , Estados Unidos/epidemiologiaRESUMO
Atopobium vaginae, a fastidious, anaerobic, Gram-positive cocci-shaped bacterium that generates large quantities of lactic acid, is associated with bacterial vaginosis (BV). Published nucleic acid amplification tests for identifying A. vaginae are directed toward the 16S ribosomal DNA with suboptimal specificity and require isolation of the organism. Here, sequencing of an A. vaginae genomic library has led to the development of a highly specific and sensitive real-time PCR test for detection of A. vaginae directly from gynecological cervicovaginal swab samples. The real-time PCR did not cross-react with DNA extracted from other members of the Atopobium genus, species with closely related 16S ribosomal DNA, and a panel of 51 other human pathogens. The DNA extraction and PCR assembly were amenable to automation using Corbett Robotics X-tractor Gene and CAS-4200N liquid handling systems. The real-time PCR was used to analyze 96 cervicovaginal swab samples submitted to our clinical laboratory for detection of organisms associated with BV. Of those samples, 28 were positive for A. vaginae. Of the 28 positive samples, 23 were concomitant with Gardnerella vaginalis detection. These results suggest that further clinical study of the relationship of A. vaginae with G. vaginalis and the development of BV should be performed.
