Development of multiplex-PCR assay for rapid detection of <em>Candida spp.</em>

• Main Authors: Ni Made A. Tarini, Mardiastuti H. Wahid, Fera Ibrahim, Andi Yasmon, Samsuridjal Djauzi
• Format: Article
• Language: English
• Published: Faculty of Medicine Universitas Indonesia 2010-05-01
• Series: Medical Journal of Indonesia
• Online Access: http://mji.ui.ac.id/journal/index.php/mji/article/view/387
• ISSN: 0853-1773; 2252-8083

<p><strong>Aim</strong> <em>Candida spp.</em> infection commonly occur in immunocompromised patients. Biochemical assay for identification of <em>Candida spp.</em> is time-consuming and shows many undetermined results. Specific detection for antibody, antigen and metabolites of <em>Candida spp.</em> had low sensitivity and specificity. In this study, we developed a rapid diagnostic method, Multiplex-PCR, to identify <em>Candida spp.</em></p><p><strong>Methods</strong> Five <em>Candida spp.</em> isolates were cultured, identifi ed 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.</p><p><strong>Results</strong> DNA detection limit by Multiplex-PCR assays for <em>C. albicans, C. tropicalis, C. parapsilosis, C. krusei</em> and <em>C. glabrata</em> 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.</p><p><strong>Conclusion</strong> Multiplex-PCR is much more sensitive than culture and thus, can be recommended as a sensitive and specific assay for identification of <em>Candida spp.</em> <em><strong>(Med J Indones 2010; 19:83-7)</strong></em></p><p><strong>Keywords:</strong> <em>Candida spp., multiplex-PCR</em></p>