Development of multiplex-PCR assay for rapid detection of Candida spp.
Aim Candida spp. infection commonly occur in immunocompromised patients. Biochemical assay for identification of Candida spp. is time-consuming and shows many undetermined results. Specific detection for antibody, antigen and metabolites of Candida spp. had low sensitivity and specificity. In this study, we developed a rapid diagnostic method, Multiplex-PCR, to identify Candida spp.
Methods Five Candida spp. isolates were cultured, identified with germ tube and API®20 C AUX (BioMerieux®SA) kit. Furthermore, DNA was purified by QIAamp DNA mini (Qiagen®) kit for Multiplex-PCR assay.
Results DNA detection limit by Multiplex-PCR assays for C. albicans, C. tropicalis, C. parapsilosis, C. krusei and C. glabrata were 4 pg, 0.98 pg, 0.98 pg, 0.5 pg and 16 pg respectively. This assay was also more sensitive than culture in that Multiplex-PCR could detect 2.6-2.9 x 100 CFU/ml, whereas culture 2.6-2.9 x 102 CFU/ml.
Conclusion Multiplex-PCR is much more sensitive than culture and thus, can be recommended as a sensitive andÂ specific assay for identification of Candida spp. (Med J Indones 2010; 19:83-7)
Copyright (c) 2010 Ni Made A. Tarini, Mardiastuti H. Wahid, Fera Ibrahim, Andi Yasmon, Samsuridjal Djauzi
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