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題 名 | 利用PCR和DNA探針之專一性引子鑑定Candida Albicans=Identification of Candida Albicans by Specific Primers of Polymerase Chain Reaction and DNA Probes |
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作 者 | 黃錦城; 蔡維鐘; 許瑞祥; 王西華; | 書刊名 | 中華民國微生物及免疫學雜誌 |
卷 期 | 30:1 1997.02[民86.02] |
頁 次 | 頁18-31 |
分類號 | 368.87 |
關鍵詞 | 聚合酶連鎖反應; DNA探針; 內轉錄區; 酵母菌鑑定; |
語 文 | 中文(Chinese) |
中文摘要 | Candida albicans為病原性酵母菌,以核醣體DNA上之普遍性引子,藉PCR增殖其內轉錄區(ITSI和ITSII),由於Candida屬內的ITSI和ITSII的DNA片段長度與Candida albicans相近,故無法直接作為Candida albicans之鑑定,所以必須再將PCR產物純化後以毛地黃素(digoxigenin)標識後作為DNA探針,才能與目標DNA雜交。由10種Zygosaccharomyces和8種Candida中,總共47株菌所抽取的DNA,與DNA探針雜交後,能夠呈色才是Candida albicans。另一種鑑定和檢測方法是由ITSI和ITSII之DNA序列分別找出一對專一性引子CA1、CA2 和 CA3、CA4,藉由PCR增殖法,只有Candida albicans才能被增殖出來,此種PCR增殖法之靈敏度為:純菌C. albicans之DNA量為0.1ng,約相當於10��-10�晜茞茩M,混合其他二種等量的酵母菌時,則需1ng,約相當於10��-10�穩茞茩M。 |
英文摘要 | Candida albicans is a pathogenic yeast. Two sets of universal primers were used for specific identification of Candida albicans with PCR-amplified ribosomal DNA internal transcribed spacer (ITS). Among the species of Candida, the amplitied ITSI and ITSII of DNA fragments were similar in size.fragments w ere similar in size. the PCR product was pruified and labeled with digoxigenin and used as DNA probein the detection with target DNA of Candida albicans by hybridization. Two sets of specific primers (CA1 and CA2 to amplify ITSI, CA3 and CA4 to amplify ITSSI) were designed by alignment of ribosomal ITS s equence of pathogenic Candida albicans with other species to detect C. al bicans by PCR. The sensitivity of PCR using the specific primers to detect pure culture of C. albicans was 0.1 ng (about 10��-10�� cells.)If the yeast cel ls were mixed with two other strains, there was a 10-fold decrease in sensit ivity (1ng or 10��-10�� cells) under the same PCR conditions. |
本系統中英文摘要資訊取自各篇刊載內容。