Online ISSN: 2515-8260

Keywords : PCR-RFLP


A COMPARISON OF HICROM AGAR AND PCR - RFLP FOR THE IDENTIFICATION OF CANDIDA SPECIES

Saher Khan,Dr. Mastan Singh, Dr. Vineeta Khare

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

Study Of Genetic Polymorphism In Oxidative Stress Related Genes And Their Association With Gastrointestinal Cancer: A Case Control Study From Rural Population Of South Western Maharashtra

Madhavi N. Patil; Anand Gudur; Rashmi Gudur; Satish Kakade; Sandeep Kadam; Suraj Pawar; Kailas D. Datkhile

European Journal of Molecular & Clinical Medicine, 2020, Volume 7, Issue 10, Pages 42-52

Oxidative stress is known to be one of the major factors involved in the development and progression of cancer. Superoxide dismutase and Catalase plays an important role in the primary defence against oxidative stress. The present study was therefore undertaken to investigate the association between polymorphism of superoxide dismutase (SOD1, SOD2, SOD3) and Catalase (CAT1 & CAT2) and the gastro intestinal cancer risk in the rural population of the south western Maharashtra. The present case-control study included 200 confirmed gastrointestinal (GI) cancer patients and 400 age and gender matched healthy controls. The polymorphism in the Superoxide Dismutase and Catalase genes was studied out by PCR-RFLP method. When we studied the genotypic frequency of SOD1, SOD2, & SOD3, SOD2 (rs1141718) showed the negative association with the GI cancer risk [OR: 0.3097, CI: 0.1727-0.5553, p <0.0001]. There was no significant association found between SOD1 & SOD3 of superoxide dismutase and CAT1 & CAT2 of Catalase gene polymorphism and susceptibility of GI cancer. The present study shows no significant association of SOD1 (G allele of rs 2070424), SOD3 (A allele of rs2536512), CAT1 (T allele of rs7943316) and CAT2 (rs1001179) with the development of gastrointestinal cancer in the rural population of south western Maharashtra from India. However, SOD2 (rs1141718) shows the negative association with the development of GI cancer.