摘要
为比较5种弓形虫抗原的诊断效果,建立实用的羊弓形虫病诊断方法,本研究制备了弓形虫速殖子全虫抗原和4个重组蛋白(GRA1、MIC3、MAG、BAG1),正交试验比较其免疫反应性,从中筛选出相对优势抗原并建立羊弓形虫病间接ELISA(iELISA)诊断方法,用于检测弓形虫特异的IgG抗体。研究结果表明:除BAG1外上述4种抗原均具有较好的免疫反应性,综合考虑选择致密颗粒蛋白1(GRA1)作为诊断抗原建立了GRA1-iELISA,其检测弓形虫抗体灵敏度大于1∶100(血清稀释度),且不与羊莫氏巴贝斯虫、吕氏泰勒虫、捻转血毛线虫阳性血清发生交叉反应,特异性良好;运用建立的方法和改良凝集试验分别对采自山羊屠宰场的80份血清样品进行检测,测得的弓形虫感染阳性率分别为28.75%(23/80)和31.25%(25/80),二者总符合率为82.5%。本研究建立的ELISA检测方法对羊弓形虫病的诊断以及商业化试剂盒的开发具有重要参考价值。
In order to develop reliable methods for the diagnosis of ovine toxoplasmosis,the whole lysates of Toxoplasma tachyzoites and four recombinant proteins(GRA1/MIC3/MAG/BAG1)were prepared for testing their suitability in indirect-ELISA.The results showed that other four antigens except BAG1 had satisfactory immunoreactivity with goat antibodies.As a fact,the recombinant protein GRA1 was selected as the optimal antigen for this purpose.The GRA1-iELISA method was optimized for serum dilution at 1:100.Additionally,GRA1-iELISA had good specifi city as it did not have cross reaction with Babesia motasi,Theileria luwenshuni and Haemonchus contortus positive sera.Finally,GRA1-iELISA and modifi ed agglutination test(MAT)were used to test 80 serum samples collected from a goat slaughterhouse.The positive rates were 28.75%(23/80)for GRA1-iELISA and 31.25%(25/80)for MAT and these two methods had a good coincidence rate at 82.5%.Overall,the GRA1-iELISA method developed in this study provided an important insight into the design of diagnostic tools for ovine toxoplasmosis.
作者
李迎迎
侯伦
王敏
李亚琼
李明俊
周艳琴
赵俊龙
申邦
LI Lingying;HOU Lun;WANG Min;LI Yaqiong;LI Mingjun;ZHOU Yanqin;ZHAO Junlong;SHEN Bang(State Key laboratory of Agricultural Microbiology,College of Veterinary Medicine,Huazhong Agriculture University,Wuhan 430070,China;Key laboratory of Veterinary Medicine in hubei province,Wuhan 430070,China)
出处
《中国动物传染病学报》
CAS
北大核心
2022年第1期127-133,共7页
Chinese Journal of Animal Infectious Diseases
基金
国家重点研发计划(2017YFD0501304)
中国农业科学院兰州兽医研究所(平台-TDRC-22)。
