Frequency And Species Of Oral Candida In Sjogren Syndrome And Genotype Distribution Of Candida Ablicans

[Objective]Compare the oral clinical isolates yeast frequency and species and C. ablicans genotype distribution between sjogren syndrome and healthy people.Discuss the Pathogenic mechanism and the clonize of clinical isolates candida strains. Find the prevalent and invasive candida spp.and the invasive genetype of Candida ablicans.Then supply the full basic research basis for preventing the oral candidiasis and developing the treatment new and effective candida yeast infections drugs.[Methods] Mucosal swab samplings were taken from46sjogren syndrome and332negative subjects in the First Affiliated Hospital of Kunming Medical University by simple cluster sampling. Microscope microscopic inspecteons discriminate and eliminate the positive one then plated onto Sabouraud's dextrose agar (SDA), incubating at37℃for48h. The isolates were identified and purified by CHROMagar Candida agar medium, and API20C AUX system for yeasts. Extracte the DNA of C. ablicans by DNA extraction kit. Then Use the25SrDNA intron part specific primer CA-INT for geneotype of C. ablicans by polymerase chain reaction (PCR).The purification PCR products A and B type were digested by MspⅠ and HaeⅢ overnight,37℃of restriction fragment length polymorphism (RFLP) method for subgenotype. Analysising the DNA sequencing directly of different genotype, find the homologous and difference.With17.0SPSS software related data analysis.Then analyse the high prevent factor of oral candida in sjogren syndrome by genotyping analysis of C. ablicans and sequence analysis.[Results] 1.A total of48female with a median age of56.5±10yearsand332healthy female with mean age of46.8±10years were enrolled in the study. Among the48sjogren syndrome,37of them77.1%(37/48) were isolated candida yeast.67of healthy people20.2%(67/332) were colonized by candida strains. The flora of clinic isolated oral candida in sjogren syndrome were much higher than the healthy.(P<0.05).2.The isolated oral candida average colony-forming unit(CFU) of sjogren syndrome is194.9±55CFU/mLwhen the healthy people is30.6±15CFU/mL.It obvious that sjogren syndrome was significantly higher than the healthy people(P<0.05).3.A total of41oral candida isolates were clonized in sjogren syndrome. The isolates include the most prevalent C.albicans strains which is32.nealy78%(32/41). Non-C.albicans individual include virous Species include7C. tropicalis about17.1%(7/41),1C. glabrata and1C. guilliermondii nearly2.4%.(1/41). A total of87oral candida isolates were clonized in healthy people. The isolates include the most prevalent C.albicans strains which is53,nealy60.9%(53/87),Non-C.albicans individual include virous Species.the most prevalent is10C. parapsilosis,11-5%(10/87).6C. glabrata6.9%(6/87) and5C.tropicalis nealy5.7%(5/87),3C. guilliermondii and3C.krusei3.4%(3/87),2C. collculosa2.2%(2/89).there were only1isolates inculde C. famata,C.lusitaniae,C. boidini,Iricho mucoides, Rnodotorule guitins. The frequency of C. ablicans, C. glabrata and C. guilliermondii isolates between sjogren syndrome and the healthy people has no significantly different(P>0.05),but sjogren syndrome were much higher than the healthy people in C. tropicali(P<0.05).The reaserch found C.krusei. C. parapsilosis,C. collculosa et al that these8species only positive in healthy people.4.There were4multiple species symbiose in sjogren syndrome nearly10.8%.the frequently islolates were C.albicans+C. glabrata. A total18isolates were multiple symbiose in healthy people.,16of them were2specie oexisted.the other2strains has3spp. The healthy group engraftment candida mycoderma slightly higher than the proportion of sjogren syndrome, but no significant differences(P>0.05).5.A total of32oral candida isolated,including20genotype A.7genotype B and5 genotype C of C. albicans in sjogren syndrome when the healthy people isolated20genotype A,14genotype B and19genotype C of C. albicans. The two groups were not isolated genotype D and E. Genotype A sjogren syndrome group were up sharply from the healthy crowd (P<0.05). Type B, two groups of no difference (P>0.05). Genotype C, health group were higher than sjogren syndrome (P<0.05).6.PCR products of20genotype A C.albicans isolates in sjogren syndrome,were digested with Mspl and HaeⅢrestriction endonucleases for5subtypes, a subtype (55%), b subtype2(10%), c subtypesl(5%), d subtypes4(20%), f subtype2(10%). PCR products of20genotype A C.albicans isolates in heathy people were digested with MspⅠ and HaeⅢrestriction endonucleases for6subtype, a subtype4(20%), b subtype2(10%), c subtype1(5%), e subtypes9(45%), g subtype2(10%), h subtype2(10%). Two groups of comparisons a subtype of sjogren syndrome is more frenqunce than health group(P<0.05), b and c subtypes were no difference. Sjogren's syndrome isolated d, f subtypes in healthy people did not appear and healthy people from g, h in Sjogren's syndrome were not isolated.7.A11of RFLP products for B were the same subtypes as (251,110,93,74,30bp) and (540,190,150)[Conclusion]1.The oral candida colonization rate and colony-forming unit in the sjogren syndrome was higher than that in heathy people.2.1n sjogren syndrome grounp, C. albicans is the predominant species. The non-C. albicans species predominantly consisted of C. tropical, which were characterized by pathogenicity and high resistance rate of azole antifungal agents. C. parapsilosis was predominant specie of non-C. albicans fungal species from normal subjects, which was short-term colonization and not obvious pathogenicity.3.The candida spp.distribution of healthy people is more than sjogren syndrome. but the symbiosis phenomena of candida between two groups has no significant differences.4. A were the primary geneotype in sjogren syndrome,is obviously higher than the healthy people in which the most common were C genetype,but A,B,C distributed relatively uniformity.5.8subtype of C ablicans's A genetype showed sequence simililarities ranging between78%-96.5%.Subtype a carried95.2%sequence simililarity to subtype e, Sequence mutations concentrate in112,113and126sites. special subtypes d,f of SS carried94.6%,93.8%sequence simililarity to h subtypes of healthy people.6. All of RFLP products for B were the same.