RESUMEN
Candidemia is one of the most common fungal nosocomial infections worldwide. It causes high mortality and morbidity rate with significant hospital costs due to increased length of hospital stay and costs for anti-fungal treatment. This study aims to investigate anti-fungal drug susceptibility, enzymatic activity and biofilm formation of the Candida spp. isolated from blood cultures. In 2016, a total of 84 clinical Candida isolates were analyzed for minimum inhibitory concentration (MIC) against fluconazole and amphotericin B by agar diffusion E-test (E-strips). Three enzymatic activity tests for phospholipase, proteinase and esterase were performed by using egg yolk agar, bovine serum albumin medium and Tween 80 opacity medium, respectively. Biofilm formation was determined by crystal violet staining. To describing the various Candida distributions cultured, C. albicans was the most frequent species (n=37, 44.1%), followed by C. tropicalis (n=30, 35.7%), C. parapsilosis (n=8, 9.5%), C. glabrata (n=6, 7.1%) and C. guilliermondii (n=3, 3.6%). Regarding anti-fungal drug susceptibility, C. albicans was susceptible to fluconazole (100%). In addition, all clinical Candida isolates were fully susceptible to amphotericin B (100%). The predominant enzyme activity of C. albicans included medium to high levels of phospholipase, proteinase and esterase activities. C. tropicalis displayed esterase activity, while C. glabrata and C. guilliermondii had no phospholipase and proteinase activity. Non-albicans Candida (NAC) i.e. C. tropicalis formed a biofilm at a higher rate than C. albicans. This study revealed the production of virulent factors in Candida strains from candidemia patients.
Asunto(s)
Antifúngicos/farmacología , Cultivo de Sangre , Candida/efectos de los fármacos , Candida/patogenicidad , Candidemia/microbiología , Biopelículas/crecimiento & desarrollo , Candida/enzimología , Infección Hospitalaria/microbiología , Esterasas/metabolismo , Humanos , Pruebas de Sensibilidad Microbiana , Péptido Hidrolasas/metabolismo , Fosfolipasas/metabolismo , Factores de VirulenciaRESUMEN
AIMS: To exploit immunomagnetic separation combined with PCR with confronting two-pair primers (IMS-PCR-CTPP) as a rapid method for detection of Mycobacterium tuberculosis complex (MTC) and identification of Mycobacterium bovis from sputum specimens. METHODS AND RESULTS: Monoclonal antibody (mAb) against the mycobacterial antigen, 85B (Ag85B), was coupled with magnetic particles for specific immunomagnetic separation (IMS) of Mycobacterium spp. Immunofluorescence assay indicated the capability of mAb to bind to Ag85B in both the recombinant and the native form. The IMS combined with PCR-CTPP targeting the mycobacterial lep B gene was further implemented using 133 sputum samples with acid-fast bacilli grading from negative to 3+. The results showed the sensitivity and specificity of IMS-PCR-CTPP vs gold standard culture method were 89·9 and 88·6% respectively. CONCLUSIONS: The IMS-PCR-CTPP method shortens the time for tuberculosis (TB) diagnosis from months to a day. This method is also suitable for investigation of MTC and epidemiological study of Myco. bovis in sputum specimens. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first report emphasizing the combination of IMS and PCR-CTPP for the detection of MTC and simultaneous identification of Myco. bovis from sputum. It could be used for TB diagnosis in resource-limited countries with high TB burden.