Keywords : Candida species
A COMPARISON OF HICROM AGAR AND PCR - RFLP FOR THE IDENTIFICATION OF CANDIDA SPECIES
European Journal of Molecular & Clinical Medicine,
2022, Volume 9, Issue 4, Pages 1501-1509
Aim: The study’s aim was to compare the effectiveness of HiCHROMagar and PCR-restriction fragment length polymorphism (PCR-RFLP) in identifying Candida species in order to ascertain the advantages and drawbacks of each technique.
Methods: One hundred and twenty five Candida strains were isolated from different clinical specimens from patients admitted to different wards of Era's Lucknow Medical College. The Candida isolates were from urine, Sputum, Blood, vaginal swab, pus and nail. Clinical samples werecultured on Sabouraud-dextrose agar (SDA) with chloramphenicol for 48 h at 37 °C.to obtain candida colonies. Wet mount,Gram staining, germ tube, sugar fermentation and assimilation were performed for all isolates. Candida isolates on SDA were subcultured on HiCrome Candida Differential agar (HiMedia, Mumbai, India) for 24-48 hours; plates were incubated at 37 °C and was re-evaluated by employing digestion of the ITS1–5.8SrDNA–ITS2 region using Msp I restriction enzyme for RFLP and universal primers internal transcribed spacer 1(ITS1) and ITS4 for PCR amplification.
Results: Out of 125 patients 58.4% were female and remaining was male. The mean age of the patients was 44.76 years. Most of the sample collected from the urine..Candida isolates identified as C.albicans based on colour (light green) by HiCrome agar were in agreementwith PCR-RFLP while , the identifcation of the non albicans Candida species (C. krusei, C. glabrata ,C.parapsilosis and C. tropicalis) by colour code on HiCrome agar showed a discrepancy with PCR–RFLP.
Conclusion:Hi chrome media has the benefit of being inexpensive and less challenging than other molecular methods, but it fail to clearly identify some uncommon candida species while RFLP-PCR employing ITSI and ITS4 primers and restriction enzyme is a trustworthy, and useful approach for identifying medically significant Candida spp. in clinical laboratories