Development of multiplex-PCR assay for rapid detection of Candida spp.
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Date
2010-05
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Abstract
Aim Candida spp. infection commonly occur in immunocompromised patients. Biochemical assay for identifi cation
of Candida spp. is time-consuming and shows many undetermined results. Specifi c detection for antibody, antigen and
metabolites of Candida spp. had low sensitivity and specifi city. In this study, we developed a rapid diagnostic method,
Multiplex-PCR, to identify Candida spp.
Methods Five Candida spp. isolates were cultured, identifi ed with germ tube and API® 20 C AUX (BioMerieux® SA) kit.
Furthermore, DNA was purifi ed 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 and
specifi c assay for identifi cation of Candida spp.
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Keywords
Candida spp., multiplex-PCR
Citation
Tarini Ni Made A, Wahid Mardiastuti H, Ibrahim Fera, Yasmon Andi, Djauzi Samsuridjal. Development of multiplex-PCR assay for rapid detection of Candida spp. Medical Journal of Indonesia. 2010 May; 19(2): 83-88.