- Multiplex Polymerase Chain Reaction Assay for Simultaneous Detection of Candida albicans and Candida dubliniensis
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Young-Hee Lim , Do-Hyun Lee
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J. Microbiol. 2002;40(2):146-150.
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 Abstract
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A multiplex polymerase chain reaction (PCR) assay was developed for the identification of two Candida species-albicans and dubliniensis. Three sets of primers were selected from different genomic sequences to specifically amplify a 516 bp fragment within the top2 gene, specific for several species of the genus Candida (CCL primers); a 239 bp fragment within the [alpha]INT1 gene, specific for Candida albicans (CAL primers); and a 175 bp fragment within the ALSD1 gene, specific for Candida dubliniensis (CDL primers). Using the primers in conjunction (multiplex PCR), we were able to detect both C. albicans and C. dubliniensis and to differentiate between them. The detection limit of the PCR assay was approximately 10 cells per milliliter of saline. Thus, this multiplex PCR assay can be applied for differentiation of C. albicans and C. dubliniensis from clinical specimens.
- Rapid PCR Method for Detecting Candida albicans Using Primers Derived from the Integrin-like Protein Gene [alpha]INT1 of Candida albicans
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Young-Hee Lim , Do-Hyun Lee
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J. Microbiol. 2000;38(2):105-108.
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Abstract
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Oligonucleotide primers amplifying a 344 bp fragment on the integrin-like protein alpha-INT1p gene ([alpha]INT1) of Candida albicans were synthesized for screening of C. albicans from clinicalsamples by the polymerase chain reaction (PCR). The PCR specifically amplified DNA from C. albicans and none from any other Candida, fungal, or human DNA in standard strains used here. The PCR assay showed that the primers (LH1 and LH2) were specific for 26 isolates of C. albicans from clinical samples, whereas the positive fragment, 344 bp, was not amplified from 15 clinical isolates including 14 other medically important Candida species and an isolate of Saccharomyces cerevisiae. PCR was conducted on the urine samples of 20 patients and 4 samples were C. albicans positive. The detection limit of the PCR assay for C. albicans was shown to be approximately 10 cells/ml saline. The PCR system using 344 bp [alpha]INT1 as a target is more specific and rapid than the conventional culture method, and the sensitive detection method is applicable to clinical diagnosis of C. albicans infections.
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