CAJ | 학술논문

目的鉴别安氏隐孢子虫卵囊的活力.方法取适量新鲜纯化卵囊,高温灭活.首先用碘化丙啶(propidium iodide,PI)染色法对灭活和未灭活卵囊进行染色,荧光显微镜下观察.同时提取灭活和未灭活两种卵囊的mRNA,反转录作为模板.基于隐孢子虫热激蛋白70(heat shock protein 70,HSP70)基因设计特异引物,用RT-PCR法对卵囊进行鉴别.结果灭活卵囊经P1染色在荧光显微镜下呈亮红色,而新鲜纯化卵囊无此颜色.新鲜纯化卵囊经RT-PCR法可检测到HSP70的特异条带,而灭活卵囊则无此条带.结论 P1染色法与基于HSP70 mRNA降解的RT-PCR法均可对隐孢子虫卵囊的活力进行鉴别.
목적 감별안씨은포자충란낭적활력.방법 취괄량신선순화란낭,고온멸활.수선용전화병정(propidium iodide,PI)염색법대멸활화미멸활란낭진행염색,형광현미경하관찰.동시제취멸활화미멸활량충란낭적mRNA,반전록작위모판.기우은포자충열격단백70(heat shock protein 70,HSP70)기인설계특이인물,용RT-PCR법대란낭진행감별.결과 멸활란낭경P1염색재형광현미경하정량홍색,이신선순화란낭무차안색.신선순화란낭경RT-PCR법가검측도HSP70적특이조대,이멸활란낭칙무차조대.결론 P1염색법여기우HSP70 mRNA강해적RT-PCR법균가대은포자충란낭적활력진행감별.
Objective To discriminate the viability of oocysts of Cryptosporidium andersoni.Methods Fresh oocysts were purified from cow faeces and then inactivated by high temperature as dead oocysts.To distinguish viable C.andersoni oocysts from dead ones,PI-dye assay Was conducted.At the same time,mRNA was extracted from fresh and inactivated cccysts respectively and RT-PCR was performed to observe the degradation of HSP70 mRNA.Results High temperature treated sample was dyed light-red by PI under fluorescence microscope while fresh oocysts were colorless.Specific band of HSP70 could be detected in fresh oocysts extract while no specific band was observed in inactivated oocysts by RT-PCR.Conclusion PI-dye assay and detection of HSP70 mRNA by RT-PCR Can both be used as method to discriminate the viability of C. andersoni oocysts.