| In general, Candida is a kind of opportunistic pathogenic yeasts. With the development of organ transplant, the increasment of malignant, AIDS and Zonked people, systemic infection especially Candida infection gets more and more, it has been the important cause for patients catching kinds of diseases go to death. With detection methods development, infection spectrograph of yeast is changed too. Anti-fungal used not reasonably, cause resistant yeasts rise continually, the mechanism of resistant gets rather complex.The mechanism of drug-resistant of Candida is rather complex, what's it come out of, judge from micro- biology standard, it's the result of the resistant gene express. However, in foretime study, the method using known genetic site detect resistant gene, gain isolated resistant gene, thus can't picture the Candida pathogenic yeasts spectrograph systemically.Vaginal Candidiasis is a commen disease in Gynaeology and Obstetrics. About seventy five percent inferty women catch once at least in their lives. With kinds of local medicine and oral anti-fungal drug come into using, there have got a better effect in treating vaginal Candidiasis. But with the using of anti-fungal medicine universally, reports about resistant facts is growing too, partial cases catch recurrent, became resistant cases. Although Vaginal Candidiasis won't danger patient's life like as deep fungal infection, itinfluence patients life quality obviously, and because of it's high occurrence rate, the<sub>5 <sub>influence is large. In Vaginal Candidiasis , the Candida albicans lies over eighty percent, in recent report, the resistant yeasts is growing, which main in other Candidas, such as Candida parapsilosis. Fungal culture identify in vitro bases on fungal phenotype, subjective results its direction in clinical limited.To explore the genetic discern among samilar phenotype yeasts, to observe the RNA discern between resistant and sensetive yeasts, this study based on Candida genome, with RAPD method and RAP-PCR in respectively. Random amplificated polymorph DNA (RAPD) is a new genetic analistic technology developmed on PCR. PCR founded in 1985, as a amplification technology in vitro, it acts in life science study importantly in recent ten years. In 1990 year, Willm and Welsh founded a new method in exploring genetic polymorph. The mechanism of RAPD as routine PCR. In RAPD amplification, DNA genome is amplificated in low strict condition , with a pair of random primers. This low strict condition allowed primer anneal decline to many sites in genome, gain a series of amplification DNA fragments. Using PAGE or electroporesis make polymorph analysis, because of the primer is synthesized randomly, needn't know the base's sequence in genome before, through increase random primer quantity, it can analysis any complex biology changes. The result is compared and analyzed by electroporesis, explore the differentiate among genome. ResultFungus cluster phenotype identify result Three hundred divided yeasts were surveyed divided from paients , there were analyzed by meli'a fungus identify system, and checked out six fungus strains all. They were Candida albicans, Torulopsis glabrata, Candida parapsilosis, Candida tropcalis, Candida krusei and Vintage barm. Using ATB FUNGUS drug sensitive project observing the Minimum Inhibitory Concentration(MIC). Three hundred divided fungi yeasts were faced to six anti-fungus medicine, which wereFlucytosin, Econazole, Ketoconazole, Miconazole, Amphotericin B and Nystatium. Finally, one hundred fifty four yeasts were choosen into study.RAPD-PCRChoice of primer In our principium test, four primers were filtered, thereinto, primer ERIC I and ERIC II can show the polymorph among Candida strains in RAPD-PCR respond. Using ERIC primers amplifiering one hundred fifty four typical sensitive and resistant yeasts, there gain stable RAPD spectrum. Spectrum amount range from four to eight. The biggest fragment exceed seven hundred bp and the smallest fragment less than one hundre... |
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