不同提取方法对食蟹猴粪便中结肠小袋虫PCR检测的影响  

Influence of Different Extraction Methods on PCR Detection of Balantioides coli in Machin

作　　者：贺会利 谢永平[1,2] 庞彬辉 毛素梅[3] 曾飞 HE Hui-li;XIE Yong-ping;PANG Bin-hui;MAO Su-mei;ZENG Fei(Guangxi Veterinary Research Institute,Naning,Guangxi,530001,China;Guangxi Key Laboratory of Veterinary Biotechnology,Nanning,Guangxi,530001,China;Guangxi Guidong Quadrumana Development & Laboratory Co.,Ltd,Wuzhou,Guangxi,543100,China)

机构地区：[1]广西兽医研究所,广西南宁530001 [2]广西兽医生物技术重点实验室,广西南宁530001 [3]广西桂东灵长类开发实验有限公司,广西梧州543100

出　　处：《动物医学进展》2021年第1期33-36,共4页Progress In Veterinary Medicine

基　　金：广西科学研究与技术开发项目(桂科能AD17129024,桂科AD20159087)。

摘　　要：比较不同抽提方法对结肠小袋虫DNA提取效率的影响,从而获得一种高效、稳定的提取粪便样品总DNA的方法,继而为后续试验提供基础材料。采用3种商品化基因组DNA提取试剂盒和经典酚-氯仿法,分别对食蟹猴粪便样品进行DNA提取,作为模板,进行PCR检测,观察不同抽提方法对DNA扩增效果的影响。4种提取方法均能扩增出目的条带,粪便抽提试剂盒的PCR扩增效果最好,测定DNA含量达到42.05 ng/μL。同时利用粪便抽提试剂盒对广西地区采集的68份食蟹猴粪便样品进行DNA提取,经PCR检测其阳性率为17.65%(12/68)。4种提取方法中以粪便抽提试剂盒对粪便样品DNA提取后经PCR扩增效果最好,可为临床粪便样品DNA提取PCR检测提供参考。In order to choose an effective and stable method from feces samples in the subsequent experiments,we compared the effect of different extraction methods on the DNA extracted efficiency of Balantioides coli.Three genomic DNA extraction kits and the classical phenol-chloroform extraction method were used to extract the DNAs from machin feces,the extracted genomic DNA was used as templates for PCR detection and the amplification results were compared.The four extraction methods can be used for amplifiying the target fragments,the best amplification effect of fecal extraction kit,which the DNA content can reach 42.05 ng/μL.The DNAs of 68 machin feces samples which collected in Guangxi with fecal extraction kit,the positive rate was 17.65%(12/68)by PCR assay.Fecal extraction kit has the best amplification from the four extraction methods.It provides important reference value for PCR detection of DNA extraction from clinical fecal samples.

关 键 词：结肠小袋虫 粪便样品 聚合酶链反应检测 脱氧核糖核酸提取 

分 类 号：S852.724[农业科学—基础兽医学]