發(fā)布時間：2018-06-17 02:49

  本文選題：申克氏孢子絲菌 + 孢子絲菌病��； 參考：《大連醫(yī)科大學(xué)》2005年碩士論文

【摘要】：申克氏孢子絲菌是孢子絲菌病的致病菌,主要引起皮膚、皮下組織和附近淋巴系統(tǒng)的亞急性和慢性感染,偶可播散至骨骼和內(nèi)臟,甚至發(fā)生全身播散性感染。真菌傳統(tǒng)的分型方法主要依賴菌體培養(yǎng)和鏡下的形態(tài)學(xué)觀察,而申克氏孢子絲菌具有“表型特征基本一致”的特點,此時,利用分子生物學(xué)方法對其進(jìn)行基因分型就顯示出一定的優(yōu)勢。本研究采用核糖體基因限制性內(nèi)切酶多態(tài)性分析(rDNA RFLP)與Southern blotting 雜交相結(jié)合的方法對孢子絲菌進(jìn)行種內(nèi)分型, 探討孢子絲菌的基因?qū)W特征,研究基因分型與菌種地區(qū)來源及臨床表現(xiàn)的關(guān)系。同時應(yīng)用常規(guī)真菌學(xué)和PCR-序列分析方法對其中一株致皮膚播散型孢子絲菌病的臨床分離株(Dmu11)進(jìn)行鑒定,并探討該菌株與皮膚淋巴管型、固定型孢子絲菌菌株在基因水平上的異同。 材料和方法:1.31 株來源于不同地區(qū)及不同臨床類型的申克氏孢子絲菌臨床分離株。CTAB 法提取菌體基因組DNA;以真菌通用引物ITS4[5‘TCCTC CGCTT ATTGA TATGC3’] 、NS5[5‘AACTT AAAGG AATTG ACGGA AG 3’]擴增ATCC10268 的rDNA 序列作為探針,與經(jīng)限制性內(nèi)切酶ApaI完全酶切后的31 株孢子絲菌DNA 進(jìn)行印記雜交。以雜交帶型作為對孢子絲菌進(jìn)行基因分型的依據(jù)。2. 常規(guī)真菌學(xué)鑒定和使用PCR-序列分析方法擴增一株致皮膚播散型孢子絲菌病臨床分離株(Dmu11)的核糖體保守區(qū)(5.8SrDNA,ITSⅡ)基因、非轉(zhuǎn)錄區(qū)(NTS)基
[Abstract]:Sporocystis schenckii is a pathogen of spore filariasis, which mainly causes subacute and chronic infections in skin, subcutaneous tissue and adjacent lymphatic system, occasionally spreading to bone and viscera, and even to systemic disseminated infection. The traditional classification methods of fungi mainly depend on cell culture and morphological observation under microscope, while Sporocystis schenckii has the characteristics of "basically consistent phenotypic characteristics", at this time, Using molecular biological methods to genotyping them shows certain advantages. In this study, ribosomal restriction endonuclease polymorphism analysis and Southern blotting hybridization were used to study the genetic characteristics of spore filaments. To study the relationship between genotyping and the origin and clinical manifestations of bacteria. At the same time, routine mycology and PCR- sequence analysis were used to identify one of the clinical isolates of dermatogenic sporocystis, Dmu11, and to explore the relationship between the strain and the lymphatic type of skin. The differences and similarities in the gene level of the strains of Sporocystis immobilized. Materials and methods Genomic DNA was extracted from clinical isolates of Sporocystis schenckii from different regions and different clinical types. CTAB method was used to extract genomic DNA, and NS5 [5AACTT AAAGG AATTG ACGGA AG3'] was expanded with the universal primer ITS4 (TCCTC CGCTT ATTGA TATGC3'). The rDNA sequence of ATCC10268 was used as a probe. The DNA of 31 strains of spore filaments digested by restriction endonuclease ApaI was hybridized with imprinting. The hybridization band type was used as the basis for genotyping of spore filaments. Routine mycological identification and PCR- sequence analysis to amplify the ribosomal conserved region of its 鈪，

本文編號：2029303