TY - JOUR
T1 - Development of multiplex-PCR assay for rapid detection of Candida spp.
AU - Tarini, Ni Made A.
AU - Wahid, Mardiastuti H
AU - Ibrahim, R. Fera
AU - Yasmon, Andi
AU - Djauzi, Samsuridjal
N1 - Publisher Copyright:
© 2010, Faculty of Medicine, Universitas Indonesia. All rights reserved.
PY - 2010/5
Y1 - 2010/5
N2 - 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 sens tive and specific assay for identification of Candida spp.
AB - 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 sens tive and specific assay for identification of Candida spp.
KW - Candida spp.
KW - Multiplex-PCR
UR - http://www.scopus.com/inward/record.url?scp=85008881640&partnerID=8YFLogxK
U2 - 10.13181/mji.v19i2.387
DO - 10.13181/mji.v19i2.387
M3 - Article
AN - SCOPUS:85008881640
SN - 0853-1773
VL - 19
SP - 83
EP - 87
JO - Medical Journal of Indonesia
JF - Medical Journal of Indonesia
IS - 2
ER -