Detection of Candida Species in Saliva of Type 2 Diabetic and Non-Diabetic Individuals using Chrom Agar Media and PCR: A Comparative study

Abilasha, J V (2020) Detection of Candida Species in Saliva of Type 2 Diabetic and Non-Diabetic Individuals using Chrom Agar Media and PCR: A Comparative study. Masters thesis, Sree Mookambika Institute of Denal Sciences, Kulasekharam.


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BACKGROUND: Oral candidiasis is an opportunistic infection of the oral cavity. It can be a mark of systemic disease, such as diabetes mellitus and is a common problem among the immune-compromised. It is caused by overgrowth or infection of the oral cavity by yeast-like fungus, candida. The important ones are C. albicans, C. tropicalis, C. glabrata, C. pseudotropicalis, C. guillierimondii, C. krusei, C. lusitaniae, C. parapsilosis, and C. stellatoidea. C. albicans represents more than 80% of isolates from clinical infection. Over the past decade, the incidence of non albicans candida species has dramatically increased. Recently, an increasing number of publications have described populations with increased incidences of C. glabrata and their resistance to commonly used antifungal agents. AIM AND OBJECTIVES: This study aims to identify and compare different candida species in saliva of Type 2 diabetic patients and non-diabetic individuals. The objectives are to identify the candida species in saliva of type 2 diabetic patients and healthy individuals using chrom agar media and to confirm the identified candida species using PCR. This is followed by quantification of the colonies of different candida species in both diabetic and healthy individuals and its correlation. MATERIALS AND METHODS: A total of 200 participants in the age group of 45-64 years were included in the study. 100 were uncontrolled type 2 diabetic patients and 100 normal healthy individuals. After collection the saliva sample was concentrated by centrifugation at 5000 rpm for five minutes. The pellet which remained at the bottom of the tube was streaked onto chrom agar media plates. These plates were then incubated at 37°C for 3-4 days. Candida speciation is done based on the different colored colonies appearing on chrom agar culture media. Total numbers of positive cultures of each species were noted in the proforma. The mean value of number of colonies of all the candida species were calculated for the two groups. These cultured species were then sent for PCR using sabouraud dextrose agar slants for species confirmation. METHODS: Comparison of candida species within and between the groups was done by applying the unpaired t-test. The results obtained showed that there is significantly increased prevalence of candida in diabetic group compared to the control group. All the species of candida, namely, candida albicans, candida glabrata, candida parapsilosis and candida krusei showed a significantly higher (p < 0.05) occurrence in the diabetic group compared to the healthy group. The highest identified species was C. glabrata followed by C. albicans in both the groups. Also the diabetic group showed more samples with multiple species of candida compared to the healthy group. CONCLUSION: Antifungal resistance is becoming an increasingly concerning problem hence understanding the mechanisms of resistance is the key to the discovery of new treatment therapies. To date, there is a lack of understanding surrounding how C. glabrata interacts within a host and the host’s defense mechanisms while simultaneously retaining a commensal existence in areas of the body including the mouth, intestines and vaginal mucosal surfaces. We can learn much from candida glabrata, and indeed it deserves more attention, since it has an existence in a class all of its own.

Item Type: Thesis (Masters)
Additional Information: 241721301
Uncontrolled Keywords: Chrom agar, C.albicans, C. glabrata, Diabetes mellitus, Oral candidiasis
Subjects: DENTAL > Oral Pathology and Microbiology
Depositing User: Subramani R
Date Deposited: 24 Feb 2021 16:43
Last Modified: 26 Feb 2021 01:34

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