作　　者： ; ; ; ; ; ;

机构地区： 华南农业大学兽医学院

出　　处： 《中国动物传染病学报》 2010年第5期32-36,共5页

摘　　要： 环孢子虫是一类可引起人和动物腹泻的重要寄生性原虫。为建立猴源环孢子虫的巢式PCR检测方法,根据该环孢子虫18S rDNA基因序列,设计一对保守引物N18SA/N18SS和一对特异性引物CYJS/CYJA,通过阳性质粒摸索该检测方法的最佳反应条件,进行特异性和敏感性试验,并应用该方法对87份猴粪样本进行了临床检测。结果显示该巢式PCR能特异性地扩增出猴源环孢子虫目的片段,与微小隐孢子虫、阿米巴原虫等8种DNA无交叉反应,最低能检测0.2 fg的阳性质粒DNA,对临床样本的检出率比传统方法（饱和蔗糖漂浮法和改良抗酸染色法）提高2.3%~3.4%。可见,该巢式PCR方法具有较高的特异性和敏感性,对于环孢子虫病的诊断和分子流行病学调查具有重要的应用价值。 A part of 18S rDNA sequence of the protozoan was chosen as the target DNA to design one pair of species-specific primers（CYJS/CYJA） and one pair of conserved primers（N18SA/N18SS） in order to establish a nested PCR method for detection of Cyclospora sp.from monkey.The nested PCR amplification conditions were optimized.A series of tests were conducted regarding the specificity and sensitivity.The results showed that specific band of 270bp was amplified from Cyclospora sp.of monkey.The detection limit was 0.2fg of DNA.Eighty seven fecal samples from monkeys in Guangzhou were detected with both the nested-PCR and other two routine methods（saturated sucrose flotation and modified acid-fast stain）.The positive rate of the nested-PCR increased by 2.3% and 3.4% compared with that of the two routine methods.The nested-PCR reported here detects Cyclospora sp.accurately and sensitively and can be used for molecular epidemiology and diagnosis of monkey cyclosporosis in the future.

关 键 词： 猴 环孢子虫 巢式 临床检测

领　　域： [农业科学] [农业科学] [农业科学] [生物学]