Effect Of “White-Gray-Opaque’’tristable Phenotypic Switching On Virulence Activity Of Candida Albicans From VVC Patients Clinical Separation

Objective1.Understand the VVC patients c. albicans infection rate in the region and its in vitro drug sensitivity to azole.2.Use YPD medium、Lee,s glucose、Lee,sGlc NAC medium,、CO2 and temperature induced white, gray, opaque the three different cell phenotype.3.Use BSA agar medium to detect the Sap activity differences in white、gray、opaque.MethodsCollection in February 2015 to October 2015, the department of Dermatology VVC specimens of 252 cases patients with vaginal secretion, 154 cases were positive. By KOH microscopy, savary-gilliard medium, API20 CAUX identification system for positive specimens of species identification. Using ATB Fungus3 yeast fungal susceptibility test system Line of 64 cases c.albicans to azole in vitro drug sensitivity testing. The preliminary extraction of c.albicans genomic DNA was isolated, polymerase chain reaction(PCR) amplification, biological methods to test whether it is c.albicans.Using MTLa1,MTL ɑ 2 specific primers to select out the MTL homozygous type in c.albicans strains.First use YPD medium to observe the original phenotypic,then use temperature, the different of CO2 content, Lee ’s glucose medium and Lee’ s Glc NAC to induce the other cell phenotype.Finally,using BSA agar medium to detect the Sap activity differences in white、gray and opaque, and indirectly, reaction of phenotype transformation effects on c.Albicans,virulence activity.Results(1) The total collection of VVC specimens of 252 cases patients with vaginal secretion,positive culture results of 154 cases, the positive rate was 61.1%. 64 strains of c. albicans(41.6%), tropical candida 31 strains(20.1%), smooth candida 22 strains(14.3%), nearly smooth candida 8 strains(5.2%), candida krusei strains(3.2%), other 24 strains(15.6%).(2) The 64 strains were identified after the c.albicans in vitro drug sensitivity test results showed that c. albicans for 5- fluorine cytosine and amphotericin B has a higher sensitivity, to fluconazole, itraconazole, voriconazole irologic resistance were 39.01%,40.63% and 45.31%, respectively.There are cross resistance and multiple drug resistance phenomenon.(3) Using MTLa1, MTLɑ2 specific primers to select the MTL homozygous type of 64 c.albicans strains,display only 3 strains are homozygous type strains(F10and F48 are ɑ / ɑ,F22 is a/a), the rest of strains are a /ɑ heterozygous type.(4) In 64 strains,we have successfully induced 3 homozygous type strains to convert the phenotype,.Finally, we got three different kinds of cell phenotype,that are white, gray and opaque.(5)The three different kinds of cells,Sap activity is different, the active followed by gray, opaque, white.Conclusion(1) For the VVC patients, the region is still the main pathogenic bacteria white beads bacterium, sensitive to fluconazole strain is more, can be used as a clinical treatment of VVC first-line drug use.(2) Using different culture medium, temperature, the different content of CO2, can successfully induce MTL homozygous type strains to contuct Phenotypic transformation,.Finally we can obtian three different phenotype of cells,that are white, gray and opaque.(3)The virulence of the three different phenotype cells,that comes from VVC patients with isolated in c. albicans,is gray > opaque > white.