【摘要】 为了对杨凌地区某鸡场鸡群球虫病疑似病例进行确诊，采用卵囊分离及纯化、形态学方法、分子生物学方法及动物感染试验对病原进行分离鉴定。结果表明，采用饱和盐水溶液漂浮法对样品中的鸡球虫卵囊分离后，镜检发现大小约为22μm×19μm的未孢子化的球虫卵囊；将获得的球虫卵囊置于25 mg/mL重铬酸钾溶液中于28℃孵育48 h进行孢子化，镜检发现孢子化的卵囊具有分化明显的4个孢子囊，每个孢子囊含2个子孢子；PCR鉴定发现只有鸡柔嫩艾美耳球虫(Eimeria tenella)和毒害艾美耳球虫(Eimeria necatrix)分别出现了约300 bp和450 bp的目的条带，扩增产物序列与GenBank中相应的序列一致；动物感染试验发现孢子化的卵囊感染鸡只6 d后出现腹泻和血便症状，从腹泻和血便的鸡只粪便中再次分离到鸡球虫卵囊。更多还原

【Abstract】 In order to diagnose a suspected case of chicken coccidiosis in a chicken farm in Yangling,oocyst isolation and purification,morphological method,molecular biology method and animal infection test were used to isolate and identify the pathogens.The results showed that after coccidial occysts in the samples were isolated by saturation salt solution floating method,unsporulated oocysts with a size of about 22μm×19μm were found by microscope.The obtained unsporulated oocysts were incubated in 25mg/mL potassium dichromate solution at the 28℃to sporulate for 48h following microscope observation for sporulated oocysts.The result suggested sporulated oocysts contain 4 sporocysts of each which contain 2 sporozoints.Molecular biology identification found that there are only two target bands,one at Eimeria tenella and another at Eimeria necatrix,about 300bp and 450bp.The sequences of the amplified products are consistent with the corresponding sequences in GenBank.The animal infection test found that chickens infected with sporulated oocysts started to diarrhea and appeared hematochezia 6 days later,and Eimeria oocysts were again isolated from contents of feces of these infected chickens.更多还原