肠道菌群多样性变性梯度凝胶电泳分析法的建立

被引：14

作者：

刘健华

陈杖榴

李云

俞道进

机构：

[1] 华南农业大学兽医学院广东省兽药研制与安全评价重点实验室

[2] 华南农业大学兽医学院广东省兽药研制与安全评价重点实验室广东广州

[3] 广东广州

来源：

中国兽医科技 | 2005年 / 06期

基金：

国家自然科学基金重点项目;

关键词：

变性梯度凝胶电泳; 肠道菌群; DNA提取; 菌群多样性;

D O I：

10.16656/j.issn.1673-4696.2005.06.008

中图分类号：

S852.6 [家畜微生物学（兽医病原微生物学）];

学科分类号：

090601 ;

摘要：

采用珠磨法、冻融研磨法和酶法对粪便细菌进行裂解,提取粪便细菌DNA,并采用16SrRNAV3区引物(P3GCf、P2r)和V6V8区引物(U968GCf、L1401r)扩增细菌16SrRNA基因可变区,对变性梯度凝胶电泳(DGGE)方法用于肠道菌群多样性分析的各种条件进行了优化,建立了分析人和动物粪便菌群多样性的DGGE方法。2对引物的粪便DGGE结果都表明,珠磨法和冻融研磨法对细菌的裂解效果优于酶法;P3GCf、P2r引物的条带数明显多于U968GCf、L1401r引物;珠磨法PCRDGGE对普通拟杆菌、青春双歧杆菌和粪肠球菌的检测限分别为(6.4±0.1)×102、(6.1±0.7)×103和(7.2±0.2)×105个细菌,珠磨法对这3种细菌的检测限均低于酶法。结果表明,珠磨法和P3GCf、P2r引物更适宜用于粪便菌群多样性DGGE分析。

引用

页码：445 / 449

页数：5

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