本文關(guān)鍵詞：VVC患者臨床分離白念珠菌white-gray-opaque三穩(wěn)態(tài)轉(zhuǎn)換對(duì)其毒力活性的影響　出處：《山西醫(yī)科大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 外陰陰道念珠菌病 白念珠菌 表型轉(zhuǎn)換 Sap分泌活性

【摘要】：目的1、了解本地區(qū)VVC患者念珠菌的菌種分布情況及對(duì)唑類藥物的敏感性;2、通過(guò)YPD培養(yǎng)基、Lee,s葡糖糖培養(yǎng)基、Lee,s N-乙酰葡糖胺(Glc NAC)培養(yǎng)基、CO2及溫度來(lái)誘導(dǎo)white、gray、opaque三種不同表型細(xì)胞的產(chǎn)生。3、利用牛血清白蛋白培養(yǎng)基(BSA)檢測(cè)white、gray、opaque三種不同表型細(xì)胞分泌型天冬氨酸蛋白酶(Sap)的活性差異。方法納入2015年2月至2015年10月就診于本科門診,疑似VVC患者陰道分泌物標(biāo)本252例,并通過(guò)10%KOH鏡檢、沙堡弱培養(yǎng)基培養(yǎng)、CHROMagar念珠菌顯色培養(yǎng)、API20CAUX鑒定系統(tǒng)進(jìn)行培養(yǎng)分離鑒定,初步獲得64株白念珠菌,采用ATB Fungus3酵母菌樣真菌敏感性檢測(cè)系統(tǒng)對(duì)其進(jìn)行藥敏試驗(yàn)。提取64株白念珠菌的基因組DNA,真菌通用引物進(jìn)行PCR擴(kuò)增,分子生物學(xué)方法進(jìn)一步檢測(cè)其是否為白念珠菌。利用MTLa1、MTLɑ2特異性引物,篩選出MTL純合型菌株。先利用YPD培養(yǎng)基對(duì)其進(jìn)行表型初篩,再利用溫度、CO2含量的不同、Lee's葡萄糖和Lee's N-乙酰葡糖胺(Glc NAC)培養(yǎng)基來(lái)誘導(dǎo)其他兩種表型細(xì)胞的產(chǎn)生,最后利用BSA培養(yǎng)基檢測(cè)每株菌株三種不同表型細(xì)胞Sap活性大小,從而反應(yīng)白念珠菌的表型轉(zhuǎn)換對(duì)其毒力活性的影響。結(jié)果(1)共收集VVC患者陰道分泌物252份,真菌培養(yǎng)陽(yáng)性者154例,陽(yáng)性率為61.1%。其中白念珠菌64株(41.6%),熱帶念珠菌31株(20.1%),光滑念珠菌22株(14.3%),近平滑念珠菌8株(5.2%),克柔念珠菌5株(3.2%),其它24株(15.6%)。(2)64株白念珠菌藥敏試驗(yàn)結(jié)果:白念珠菌對(duì)5-氟胞嘧啶和兩性霉素B有較高的敏感性,對(duì)氟康唑、伊曲康唑、伏立康唑的耐藥率分別為39.01%、40.63%和45.31%,存在交叉耐藥現(xiàn)象。(3)64株菌株的MTL a、ɑ基因篩查顯示只有3株為純合型菌株(F22為a/a型,F10、F48為ɑ/ɑ型),余均為a/ɑ雜合型。(4)64株菌株中只有3株純合型菌株成功進(jìn)行了表型轉(zhuǎn)換,誘導(dǎo)出white、gray和opaque三種不同表型細(xì)胞。(5)3株菌株white、gray、opaque細(xì)胞Sap分泌活性存在差異,其活性大小依次為gray、opaque、white。結(jié)論1、本地區(qū)VVC患者所感染的致病真菌仍以白念珠菌為主,對(duì)FCA耐藥率相對(duì)較低,可作為臨床治療的一線用藥。2、利用不同的培養(yǎng)基、溫度、不同CO2濃度,可以成功誘導(dǎo)MTL純合型菌株進(jìn)行表型轉(zhuǎn)換,得到white、gray、opaque三種細(xì)胞。3、VVC患者分離出的白念珠菌中三種不同表型細(xì)胞的毒力grayopaquewhite。
[Abstract]:Objective to understand the distribution of 1 species of local VVC in Candida and sensitivity to azole drugs; 2, by YPD medium, Lee medium, s glucose, Lee, s N- acetylglucosamine (Glc NAC) medium, CO2 and temperature to induce white, gray, opaque three different the phenotype of cells produce.3 culture medium using bovine serum albumin (BSA) detection of white, gray, opaque three different types of cells secreted aspartic proteinase (Sap) activity difference. Methods included in February 2015 to October 2015 in the outpatient clinic, patients with suspected VVC vaginal secretion samples of 252 cases, and through the 10%KOH examination sand castle, weak medium, CHROMagar Candida culture, API20CAUX identification system culture isolation and identification, obtained 64 strains of Candida albicans, drug sensitivity tests were performed using ATB Fungus3 yeast like fungi susceptibility detection system from 64 strains of Candida albicans. The genomic DNA of bacteria and fungi universal primers for PCR amplification, molecular biology method to detect whether it is Candida albicans. The use of MTLa1, MTL. 2 primers were screened for MTL homozygous strains. Using YPD culture medium on the phenotypic screening, using temperature, different content of CO2, Lee's glucose Lee's and N- acetylglucosamine (Glc NAC) to induce the other two cell phenotype medium, finally using BSA medium to detect each of three strains of different size and activity of Sap cell phenotype, response phenotype of Candida albicans virulence conversion effect on its activity. Results (1) were collected in patients with vaginal VVC the secretion of 252 copies, 154 cases of fungal culture positive, the positive rate of 61.1%. among 64 strains of Candida albicans (41.6%), Candida tropicalis 31 strains (20.1%), 22 (14.3%) strains of Candida glabrata, Candida parapsilosis (5.2%), 8 strains of Candida krusei 5 strains (3.2%), the 瀹，

本文編號(hào)：1437188