CAJ | 학술논문

为了解感染不同灵长类动物的隐孢子虫种类以及与感染人的人隐孢子虫(C.hominis)之间的遗传差异,本研究采用形态学和分子生物学方法对猴源隐孢子虫进行分离和鉴定.利用常规方法分离猴粪便中的隐孢子虫卵囊,通过改良抗酸染色和荧光显微镜观察对其进行形态学鉴定;并采用PCR方法扩增其卵囊壁蛋白(cowP)基因和18S rRNA基因,扩增产物克隆至pMD-18-T载体中,对阳性克隆进行测序并作进化树分析.结果表明:抗酸染色和荧光检查结果与所报道的人隐孢子虫的结果一致.PCR扩增产物经电泳检测,明显地出现554 bp和370bp大小的片段,与预期结果一致;两种基因的序列分析结果显示该猴源隐孢子虫与C. hominis的相似性均为100%.由此可认为本次分离的隐孢子虫为C.bominis.
위료해감염불동령장류동물적은포자충충류이급여감염인적인은포자충(C.hominis)지간적유전차이,본연구채용형태학화분자생물학방법대후원은포자충진행분리화감정.이용상규방법분리후분편중적은포자충란낭,통과개량항산염색화형광현미경관찰대기진행형태학감정;병채용PCR방법확증기란낭벽단백(cowP)기인화18S rRNA기인,확증산물극륭지pMD-18-T재체중,대양성극륭진행측서병작진화수분석.결과표명:항산염색화형광검사결과여소보도적인은포자충적결과일치.PCR확증산물경전영검측,명현지출현554 bp화370bp대소적편단,여예기결과일치;량충기인적서렬분석결과현시해후원은포자충여C. hominis적상사성균위100%.유차가인위본차분리적은포자충위C.bominis.
In this study,Cryptospofidium sp.was isolated from feces of rhesus (Macaca mulatta) and characterized by modified anti-acid fast staining and fluorescence observation.The Oocysts were shown to possess same morphological characteristics as that reported for C.hominis.Two target genes of COWP and 18S rRNA were amplified by PCR,cloned and sequenced,and the evolution tree was generated by Mega software.Both fragments shared 100%sequence identities with C.hominis and their heredity distances was the nearest to that of C.hominisin evolution tree.It is concluded that the Cryptosporidium isolated from rhesus belong to C.hominis.