Diagnosis of Candida albicans: conventional diagnostic methods compared to the loop-mediated isothermal amplification...

Candida species cause a wide range of opportunistic infections in humans and animals. The detection of Candida species by conventional diagnosis methods is costly and time consuming. This study was conducted for the first time to evaluate and compare a relatively new molecular assay and the loop-mediated isothermal amplification (LAMP) technique with conventional methods for detection of Candida albicans. In this study, 70 different species of Candida identified by conventional methods were cultured on Sabouraud chloramphenicol agar medium and then the genomic DNA was extracted. The LAMP technique was performed using specific primers targeting the ITS2 gene of C. albicans. The analytical sensitivity and specificity of LAMP were measured using a tenfold serial dilution prepared from extracted DNA from standard C. albicans strain from 1 ng to 1 fg and the DNA samples of other clinical Candida species and three non-Candida yeast. Out of 70 yeast samples analyzed by LAMP technique, 24 samples (34.3%) were positive for C. albicans. Comparison of the results showed that the CHROMagar Candida and germ tube production methods are quite consistent with the LAMP technique, while the agreement amount between the results of carbohydrate assimilation and chlamydoconidia generation assays and LAMP technique was 98.5% and 72.8%, respectively. The detection limits of the LAMP assay were 10 fg of the DNA from the standard C. albicans strain. No amplification was observed in the DNA samples of other yeast species and only the DNA sample of standard C. albicans strain was amplified. Based on the results, it can be concluded that the LAMP method is as specific and precise as common diagnostic methods, but is faster, easier deployable or more sensitive. Therefore, this method can be used as a suitable complementary assay for Candida diagnosis in medical diagnostic laboratories and field conditions.