结肠小袋纤毛虫PCR-RFLP分型方法的建立  

Establishment of a PCR-RFLP typing method for Balantioides coli

作　　者：徐啊慧 冯彩彩 丰山旺 赵立卓 齐闻新 张雯 胡苏辉 王天奇[1] 闫文朝[1] XU A-hui;FENG Cai-cai;FENG Shan-wang;ZHAO Li-zhuo;QI Wen-xin;ZHANG Wen;HU Su-hui;WANG Tian-qi;YAN Wen-chao(College of Animal Science and Technology,Henan University of Science and Technology,Luoyang 471023,China)

机构地区：[1]河南科技大学动物科技学院,洛阳471023

出　　处：《中国人兽共患病学报》2024年第9期829-833,840,共6页Chinese Journal of Zoonoses

基　　金：国家自然科学基金项目(No.31772733)。

摘　　要：目的建立高效、特异的结肠小袋纤毛虫遗传亚型分析方法。方法选择限制性内切酶ApoI和PflMI对结肠小袋纤毛虫ITS1-5.8S rDNA-ITS2的PCR扩增产物进行酶切分析,建立PCR-RFLP分型方法,利用建立的PCR-RFLP方法对猪源、羊源和豚鼠源临床粪便样品进行遗传亚型分析。结果基于ApoI和PflMI的PCR-RFLP方法可以准确区分结肠小袋纤毛虫遗传变异型A和B,用PflMI可以进一步将遗传变异型B细分为B-c和B-t两个亚型。与镜检和测序结果比较,建立的PCR-RFLP方法具有良好的特异性和更高的灵敏性,不仅可以鉴定临床样品中结肠小袋纤毛虫单个亚型,而且可以鉴别单个样品中结肠小袋纤毛虫多个亚型的混合感染。结论本研究成功建立了结肠小袋纤毛虫PCR-RFLP方法,可用于结肠小袋纤毛虫遗传多态性鉴定和分子流行病学研究。This study established an efficient and specific method for type analysis of genetic variants of Balantioides coli.The restriction endonucleases Apo I and Pfl MI were selected to digest the PCR amplification products of ITS1-5.8S rDNA-ITS2 of B.coli,and to establish a PCR-RFLP typing method.The PCR-RFLP method was subsequently used to analyze the genetic variants in clinical fecal samples from pigs,sheep,and guinea pigs.The PCR-RFLP method based on Apo I and Pfl MI accurately distinguished the main A and B genetic variants of B.coli,and further divided the main B type into B-c and B-t subtypes of genetic variants with Pfl MI.Compared with the results of microscopy and sequencing,the PCR-RFLP method showed good specificity and higher sensitivity,and was able to identify not only single but also multiple variants of B.coli in a single clinical sample.This study successfully established the PCR-RFLP method for B.coli,which can be used for genetic diversity identification and molecular epidemiological studies of B.coli.

关 键 词：结肠小袋纤毛虫 PCR-RFLP分析 猪 羊 豚鼠 

分 类 号：R382[医药卫生—医学寄生虫学]