CAJ | 학술논문

目的对犬粪中的疑似环孢子虫卵囊进行形态学和分子鉴定.方法对首次在犬粪中发现的疑似环孢子虫卵囊的形状、大小、抗酸染色、孢子化和自发荧光等形态特征进行了观察.根据GenBank已发表的环孢子虫序列设计并合成3条引物,对犬源环孢子虫18SrDNA部分序列进行巢氏PCR扩增,将扩增产物纯化并克隆至载体pMD19-T,挑选阳性克隆测序并进行同源性及系统进化分析.结果该疑似环孢子虫卵囊与人的环孢子虫卵囊形态特征相似,扩增的18SrDNA片段大小为715bp,与预期目的片段一致.序列同源性和系统发育分析显示该犬源环孢子虫与环孢子虫间的相似性最高,系统进化树中处于同一分支.结论首次从犬粪中发现的疑似环孢子虫卵囊可以确定为环孢子虫.
목적 대견분중적의사배포자충란낭진행형태학화분자감정.방법 대수차재견분중발현적의사배포자충란낭적형상、대소、항산염색、포자화화자발형광등형태특정진행료관찰.근거GenBank이발표적배포자충서렬설계병합성3조인물,대견원배포자충18SrDNA부분서렬진행소씨PCR확증,장확증산물순화병극륭지재체pMD19-T,도선양성극륭측서병진행동원성급계통진화분석.결과 해의사배포자충란낭여인적배포자충란낭형태특정상사,확증적18SrDNA편단대소위715bp,여예기목적편단일치.서렬동원성화계통발육분석현시해견원배포자충여배포자충간적상사성최고,계통진화수중처우동일분지.결론 수차종견분중발현적의사배포자충란낭가이학정위배포자충.
The morphological and molecular identifications of Cyclospora-like oocysts of dogs were underwent in the present study, in which the morphological characteristics of the Cyclospora-like oocysts firstly found in the stool samples of dogs, such as shape, size, acid-fast staining,sporulation and auto-fluorescene, were observed. According to the published sequence of the rDNA gene of Cyclospora in GenBank, 3 primers were designed and were used to amplify part of the 18S rDNA gene of dog-associated Cyclospora-like organism by nested PCR.The amplicons were purified and cloned into vector pMD19T. Then, the positive clones screened were sequenced and subjected to sequence homology and phylogenetic analysis. It was found that the morphological charactertistics of the Cyclospora-like oocysts in dogs were similar to that of the human Cyclospoa oocysts and the size of-the amplified fragment of 18S rDNA was proved to be 715 bp, that was identical to that of the target fragment. Based on the results of sequence homology and phylogenetic analysis, the dog-associated Cyclospora-like organism was identified as the Cyclospora species.