Abstract
Aims: This study aimed to detect and identify Candida glabrata isolated from denture stomatitis patients by using polymerase chain reaction( PCR). Materials and Methodes :Total of forty three of oral swabs samples were obtained from patients suffering from denture stomatitis attending prosthodontic departmentCollege of Dentistry /Mosul university/ Dental teaching Hospital. Clinically ,62 isolates of Candida spp. were identified to species level by standard culture methods using Sabouraud Dextrose agar(SDA) , HiCrome™ Candida Differential Agar followed by microscopic examination, and germ tube test. DNA extraction of Candida glabrata from broth cultures was carried out ,then molecular identification with PCR using specific primers targeting phospholipase B gene (PLB) were done to confirm C. glabrata diagnosis. Results: Among 62 isolates of Candida species , the predominant type was the Candida albicans which accounted for 29(46.8%) followed by Candida glabrata 21(33.9%), Candida tropicalis 11(17.7%), finally Candida krusei accounted for only 1 (1.6%) . HiCrome™ Candida Differential Agar do not easily recognize Candida spp. Sometimes C. glabrata was falsely identified as C. parapsilosis on HiCrome™ Candida Differential Agar. The result showed that the PCR products for the specific primer gave bands on agarose gel at the position 404 bp .Conclusion : Candida glabrata is emerging as the second most spreading among the isolates. Detection of PLB gene using PCR provides a definitive target that could be used for the identification and detection of Candida glabrata. from clinical samples .
Highlights
Induced stomatitis ( DIS). ,Candida- related DS is usually caused by C. albicans [1], 1-Sampling and cultural diagnosis : The criteria for patient selection in current recent study showed that non albicans Candida study include elderly patient from both spp.is increased and reported in many studies as a genders suffering from denture stomatitis large source of infection and markedly increased wearing complete denture
The extracted DNA for all suspected isolates of C.glabrata were subjected to Polymerase Chain Reaction (PCR) for amplifying phospholipase B gene (PLB) gene .The results confirm the presence of 404bp PCR products when compare with DNA ladder,Figure [3]
The results of the current study agreed with several reports which observed that C. glabrata can be recognized from other Candida spp. by the formation of lavender to pale pink colonies on HiCromeTM Candida Differential Agar [16] in this study the colonies of C.glabrata strains exhibited significant variability when they appeared as ivory or white large colonies, this result agreed with Hospenthal (2006) [17] noted that most nonC. albicans strains are ivory, pink, lavender on HiCromeTM Candida Differential Agar. previous study conducted by Sagar et al proved that C. glabrata falsely identified as C. parapsilosis on Hicrome agar [18]
Summary
Induced stomatitis ( DIS). ,Candida- related DS is usually caused by C. albicans [1] , 1-Sampling and cultural diagnosis : The criteria for patient selection in current recent study showed that non albicans Candida study include elderly patient from both spp.is increased and reported in many studies as a genders suffering from denture stomatitis large source of infection and markedly increased wearing complete denture. Candida- related DS is usually caused by C. albicans [1] , 1-Sampling and cultural diagnosis : The criteria for patient selection in current recent study showed that non albicans Candida study include elderly patient from both spp.is increased and reported in many studies as a genders suffering from denture stomatitis large source of infection and markedly increased wearing complete denture. Candida DNA for each isolate was extracted using candida DNA preparation kit .(igenomic BYF , DNA extraction Mini Kit, iNtRON Biotechnology, South Korea) B- polymerase chain reaction (PCR) : The primers employed for the amplification PLB gene : F5'TCTCACAC TCCATTGTCTCA-3' and R 5'AGCAGGTTTACCATCAGAA-3' [14] were provided in lyophilized forms by (IDT, USA) . DNA sample were observed by using agarose gel electrophoresis (Jena Bioscience , Germany ) prepared as reported by Maniatis et al [15] . No Step 1 Polymerase activation 2 Denaturation 3 Annealing 4 Extension 5 Final Extension 6 Hold
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