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结肠小袋纤毛虫PCR-RFLP分型方法的建立OA北大核心CSTPCD

Establishment of a PCR-RFLP typing method for Balantioides coli

中文摘要英文摘要

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 ApoI and PflMI 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 ApoI and PflMI 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 sub-types of genetic variants with PflMI.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 di-versity identification and molecular epidemiological studies of B.coli.

徐啊慧;冯彩彩;丰山旺;赵立卓;齐闻新;张雯;胡苏辉;王天奇;闫文朝

河南科技大学动物科技学院,洛阳 471023

基础医学

结肠小袋纤毛虫PCR-RFLP分析豚鼠

Balantioides coliPCR-RFLP analysispigsheepguinea pig

《中国人兽共患病学报》 2024 (009)

829-833,840 / 6

Supported by the National Natural Science Foundation of China(No.31772733) 国家自然科学基金项目(No.31772733)

10.3969/j.issn.1002-2694.2024.00.128

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