MOLECULAR CHARACTERIZATION OF Candida albicans IN WOUNDS OF DIABETIC PATIENTS IN SELECTED HEALTH FACILITIES IN NORTH CENTRAL-NIGERIA

Molecular study shows that the Internal Transcribed Spacer 1 (ITS-1) gene region is a highly variable ribosomal
DNA sequence used as a primary molecular marker for identifying Candida species in clinical samples. By
exploiting ITS-1 size polymorphism, PCR-based methods was used to identify genus Candida. This study was a
cross-sectional study carried out in selected health facilities in North Central Nigeria among adult diabetic foot
ulcer patients over a period of one year. A total of 33 samples was used for the study. Specie specific identification
involved phenotypic methods such as chromogenic character on CHROMagar Candida species medium, germ
tube examination, and microscopic characteristics. Data were analysed using WHONET and SPSS. Fungal DNA
was extracted using Qiagen DNA extraction kit and identified molecularly using ITS-1 genus specific Candida
primers also focusing on detecting virulence gene such as HWP-1 and Resistance genes such as ERG-1 and MDR1
gene using PCR techniques. Results for ITS-1 gene for genus specific Candida was at bp 525bp on 100bp ladder.
All the Candida isolates harbored virulence gene on chromosomal DNA. Specifically, (75%) carried the HWP-1
gene at 572bp (P < 0.001 no significant difference). Candida albicans harbored resistant genes on chromosomal
DNA. Specifically, (45%) carried the ERG-1 gene at 510bp and no band for the MDR1 gene. The highest HWP
1 (75%) encodes a major surface adhesin for virulence, specifically mediating the binding of hyphae to host
epithelial cells. A Maximum Likelihood phylogenetic tree was constructed using the IQTREE and the generated
file was viewed. The proportion of trees (1000 bootstrap) in which the corresponding taxa clustered. Phylogenetic
analysis confirmed their genetic affiliation showing minimal intra-specie variation. Non-albicans Candida (NAC)
contained the ERG-1 gene in C. tropicalis at 380bp and C.glabrata. This study reports a concerning prevalence
of virulence and resistant genes in Candida albicans at 480bp. The genomic presence of virulence and resistant
genes underscores the importance of molecular surveillance, improved water and sanitation infrastructure and
urgency of effective antifungal strategies in combating diabetic foot ulcer infections in vulnerable populations.

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