sdj-10148

Identification and Genotyping of Candida Species Involved in Oral Candidiasis among Diabetic Patients

Shene A. Mohammed * , Khattab A. Shekhany * , Paywast J. Jalal * , Chiman H. Fattah **

*Biology Department, College of Science, University of Sulaimani, Sulaimani, Iraq.

**Anatomy Department, College of Medicine, University of Sulaimani, Sulaimani, Iraq.

Submitted: April 3, 2022, Accepted: May 23, 2022, Published: June 1, 2022.

DOI: https://doi.org/10.17656/sdj.10147

Objective: Oral candidiasis is more prevalent among diabetic patients than non-diabetics due to the factors that promote Candida oral carriage. This study aimed to isolate and identify Candida species involved in oral candidiasis of patients with diabetes mellitus in Sulaymaniyah city. 

Methods: The study was performed from August 2021 to March 2022 on two diabetic patients, Type 1 and Type 2 (n=150) and non- diabetics (n=50), as a control group. In Iraq's Kurdistan region, oral swabs were taken from 200 participants at the Sulaymaniyah governorate's Diabetic and Endocrine Center and Shar hospital. Sabouraud dextrose agar (SDA) medium was used to culture the swabs. Candida isolates were identified using HiCromeTM Candida Differential agar, then confirmed using polymerase chain reaction based on the ITS region and CHS1 gene detection. Using the CA25S and CA-INT primers, all C. Albicans isolates were genotyped based on the transposable intron in 25S rDNA. ITS1 and ITS4 primers were used to sequence the 18S region of ribosomal DNA (rDNA). Descriptive statistics were used for summaries and to describe data.      

Results: From the samples of 150 diabetes patients and 50 controls, 64 (42.6%) and 12 (24%) were positive for Candida spp. In the diabetic patients, 34 (53.1%) of the 64 isolated Candida spp. were identified as C. Albicans, while 6 (50%) of the healthy subjects had C. albicans. The genotypes A (450 bp), B (840 bp), C (450 and 840 bp) of C. Albicans and D (1040 bp) that belongs to C. dubliniensis were detected. Genotype A (54.69%) was the most frequent.  

Conclusions: This study concluded that there was a difference in the proportion of Candida spp. colonization in the oral cavity of diabetic patients compared to the healthy group; also, we found that C. Albicans with Genotype A was the most prevalent species among all other species in both groups.                                                                                     

Keywords:  Oral candidiasis, Diabetic, C. Albicans, Genotype A, Phylogenetic tree.          

Full Article - PDF                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   

References:

1. Ksiezopolska E, Gabaldón T. Evolutionary emergence of drug resistance in Candida opportunistic pathogens. Genes (Basel). 2018;9(9):461.

2. Sardi JC, Scorzoni L, Bernardi T, Fusco-Almeida AM, Giannini MM. Candida species: current epidemiology, pathogenicity, biofilm formation, natural antifungal products and new therapeutic options. J Med Microbiol. 2013;62(1):10-24.

3. Rodrigues CF, Rodrigues ME, Henriques MC. Promising alternative therapeutics for oral candidiasis. Curr Med Chem. 2019;26(14):2515- 28.

4. Quindós G, Gil-Alonso S, Marcos-Arias C, Sevillano E, Mateo E, Jauregizar N, et al. Therapeutic tools for oral candidiasis: current and new antifungal drugs. Med Oral Patol Oral Cir Bucal. 2019;24(2):e172-80.

5. Akpan A, Morgan R. Oral candidiasis. Postgrad Med J. 2002:78(922):455-59.

6. Goyal R, Jialal I. Diabetes mellitus type 2. 2018.

7. Nguyen TD, Nguyen TTH, Tran QV. The incidence of oral candidiasis in patients with diabetes mellitus: a cross-sectional study in southern Vietnam. J Crit Rev. 2020;7(4):82-6.

8. Mohammadi F, Javaheri MR, Nekoeian S, Dehghan P. Identification of Candida species in the oral cavity of diabetic patients. Curr Med Mycol. 2016;2(2):1-7.

9. Rodrigues CF, Rodrigues ME, Henriques M. Candida sp. infections in patients with diabetes mellitus. J Clin Med. 2019;8(1):76-117.

10. Alrayyes SF, Alruwaili HM, Taher IA, Elrahawy KM, Almaeen AH, Ashekhi AO, et al. Oral candidal carriage and associated risk indicators among adults in Sakaka, Saudi Arabia. BMC oral health. 2019;19(1):1-7.

11. Hani U, Shivakumar HG, Vaghela R, Osmani RA, Shrivastava A. Candidiasis: a fungal infection- current challenges and progress in prevention and treatment. Infect Disord Drug Targets. 2015;15(1):42-52.

12. Tantivitayakul P, Panpradit N, Maudcheingka T, Klaophimai A, Lapirattanakul J. Genotyping of Candida albicans and Candida dubliniensis by 25s rDNA analysis shows association with virulence attributes in oral candidiasis. Arch Oral Biol. 2019;97(1):18-24.

13. Baker M.H.A. Genotype comparisons of strains of Candida albicans from patients with vaginal candidiasis. 2012; Ph.D. diss

14. Hemaid AS, Abdelghany MM, Abdelghany TM. Isolation and identification of Candida spp. from immunocompromised patients. Bull Natl Res Cent. 2021;45(1):1-8.

15. Mehta R, Anupama SW. Evaluation of hicrome Candida differential agar for species identification of Candida isolates from various clinical samples. Int J Contemp Med Res. 2016;3(4):1219-22.

16. Kaup S, Sankarankutty J, Balasubrahmanya HV, Kulkarni S, Nirmala M. Speciation of Candida using HiCrome Candida Differential Agar. Int J Curr Microbiol App Sci. 2016;5(7):267-74.

17. Fujita SI, Senda Y, Nakaguchi S, Hashimoto T. Multiplex PCR using internal transcribed spacer 1 and 2 regions for rapid detection and identification of yeast strains. J Clin Microbiol. 2001;39(10):3617-22.

18. Barbedo LS, Figueiredo-Carvalho MH, Muniz MD, Zancopé-Oliveira RM. The identification and differentiation of the Candida parapsilosis complex species by polymerase chain reaction- restriction fragment length polymorphism of the internal transcribed spacer region of the rDNA. Mem Inst Oswaldo Cruz. 2016;111(4):267-70.

19. Baradkar VP, Mathur M, Kumar S. Hichrom Candida agar for identification of Candida species. Indian J Pathol Microbiol. 2010;53(1):93-5.

20. Charles MP, Kali A, Joseph NM. Performance of chromogenic media for Candida in rapid presumptive identification of Candida species from clinical materials. Pharmacogn Res. 2015;7(Suppl 1):S69-73.

21. Shekhany KA. Isolation and genotyping of Candida albicans involved in vaginal candidiasis among pregnant women in Sulaymaniyah and Erbil cities. Zanco J Med Sci. 2021;25(1):493-502.

22. Nimri LF, Kaplan NM, Ishaq RI. Bloodstream yeast species and in vitro antifungal susceptibility profiles. Med Sci. 2021;6(2):117-23.

23. Jordan JA. PCR identification of four medically important Candida species by using a single primer pair. J Clin Microbiol. 1994;32(12):2962-7.

24. Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987;4(4):406-25.

25. Dopazo J. Estimating errors and confidence intervals for branch lengths in phylogenetic trees by a bootstrap approach. J Mol Evol. 1994;38(3):300-4.

26. Rzhetsky A, Nei M. A simple method for estimating and testing minimum-evolution trees. Mol Biol Evol. 1992;9(5):945-67.

27. Zuckerkandl E, Pauling L. Evolutionary divergence and convergence in proteins, in Evolving genes and proteins. Elsevier. 1965;97- 166.Academic Press.

28. Kumar S, Stecher G, Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol. 2016;33(7):1870-4.

29. Vázquez‐González D, Perusquía‐Ortiz AM, Hundeiker M, Bonifaz A. Opportunistic yeast infections: candidiasis, cryptococcosis, trichosporonosis and geotrichosis. J Dtsch Dermatol Ges. 2013;11(5):381-94.

30. Zomorodian K, Kavoosi F, Pishdad GR, Mehriar P, Ebrahimi H, Bandegani A, et al. Prevalence of oral Candida colonization in patients with diabetes mellitus. J Mycol Med. 2016;26(2):103-10.

31. Chouhan S, Kallianpur S, Prabhu KT, Tijare M, Kasetty S, Gupta S. Candidal prevalence in diabetics and its species identification. Int J Appl Basic Med Res. 2019;9(1):49-54.

32. Al-Badri AS, Ali EN, Ajah HA, Ajah HA. Assessment frequency of oral fungal infection with type 2 diabetes mellitus in Iraqi patients. Acad Glo Indersci Res. 2022;3(01):16-26.

33. Abu-Elteen KH, Hamad MA, Salah SA. Prevalence of oral Candida infections in diabetic patients. Bahrain Med Bull. 2006;28(1):1-8. Vol 9(1) Mohammed et al. 53

34. da Silva-Rocha WP, Lemos VL, Svidizisnki TI, Milan EP, Chaves GM. Candida species distribution, genotyping and virulence factors of Candida albicans isolated from the oral cavity of kidney transplant recipients of two geographic regions of Brazil. BMC Oral Health. 2014;14(1):1- 9.

35. Zhu X, Dong N, Wang Q, Cheng B, Zhu L, Tao J, et al. Genotyping and antifungal susceptibility of Candida albicans clinical isolates from Yangzhou region in China. Afr J Microbiol Res. 2011;5(25):4406-10.

36. Athab AM, Rasheed MM. Detection of Candida species phenotype and genotype in diabetes mellitus patient in baquba teaching hospital. Indian J Forensic Med Toxicol. 2019;13(4):518-22.

Abstract