文档介绍：PCR技术检测棘阿米巴26S核糖体DNA
(作者:___________单位: ___________邮编: ___________)
【摘要】目的:用聚合酶链反应(PCR)方法检测棘阿米巴的26S核糖体DNA(26S rDNA,Rns),为早期诊断棘阿米巴角膜炎提供方法。方法:分离培养三株不同基因型棘阿米巴虫株,提取虫株基因组DNA,合成棘阿米巴属特异性引物(ArDNA-a),通过聚合酶链反应(PCR)扩增部分26S rDNA序列。结果:三株不同基因型棘阿米巴虫株属于两种基因型,其中Acanthamoeba sp. CJY/s2株和Acanthamoeba T4基因型,Acastellanii Neff株属于Neff基因型。PCR扩增后,Acanthamoeba Neff株的26S核糖体DNA基因片段扩增出分子量大小为126 bp 的特定扩增带,而Acanthamoeba sp. CJY/s株的26S核糖体DNA基因片段则未得到预期的扩增带。结论:用棘阿米巴属特异性引物(ArDNA-a)可以有效扩增Acanthamoeba sp. CJY/s2和Acastellanii Neff株的26S核糖体DNA基因,可以为部分棘阿米巴性角膜炎的早期检测提供新的检测手段。
【关键词】 PCR 棘阿米巴角膜炎 26S核糖体DNA
Abstract: Objective: To provide a method for early diagnosis of acanthamoeba keratitis using the polymerase chain reaction (PCR) amplificat the gene 26s-rDNA fragment for different species of acanthamoeba. Methods: Isolated culture 3 groups of different genotype acanthamoeba, which whole genome were extracted, PCR amplificated parts of gene sequence of 26s-rDNA by using acanthamoeba specificity primer (ArDNA-a). Results: In 3 different species acanthamoebas, Acanthamoeba sp. CJY/s2 and Acanthamoeba sp. CJY/s were Rns genotype T4, Acastellanii Neff was Neff. After PCR, 26s-rDNA fragment about 126 base pairs was essfully amplified by single PCR from the genotype DNA of Acanthamoeba and Acastellanii Neff, but no anticipate fragment was obtained from the genotype DNA of A