Liu Zhijie, Li Youquan, Salih Dia Eldin A, Luo Jianxun, Ahmed Jabbar S, Seitzer Ulrike, Yin Hong
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Grazing Animal Diseases MOA, Key Laboratory of Veterinary Parasitology of Gansu Province, LVRI, CAAS, Lanzhou, PR China.
Veterinary Research Institute, Khartoum, Sudan.
Vet Parasitol. 2014 Aug 29;204(3-4):139-45. doi: 10.1016/j.vetpar.2014.05.010. Epub 2014 May 17.
An enzyme-linked immunosorbent assay (ELISA) based on a recombinant Theileria uilenbergi immunodominant protein (rTuIP) was validated for detection of antibodies in 188 positive and 198 negative reference serum samples, respectively. The cut-off value was determined at 32.7% with 95% and 90% accuracy levels by two-graphic receiver-operating characteristic (TG-ROC). The equal diagnostic sensitivity (Se) and specificity (Sp) were calculated to be 98.4%. Further validation of the repeatability with positive and negative reference samples indicated the reliable performance of the assay. Monitoring the antibody dynamics of sheep experimentally infected with Theileria luwenshuni showed the efficient detection of antibody response against the pathogen at the early infection stage and up until two months post infection. Application of this assay for detection of antibody in field sera from previous unknown Theileria endemic regions in Suizhou and Guiyang showed 17.8% and 11.6% seroprevalence, respectively, and presence of the pathogen was confirmed by identification of the 18S rRNA gene in the corresponding blood of the seropositive animals. These data support that the rTuIP ELISA could be a useful tool to study the epidemiology of theileriosis caused by T. uilenbergi and/or T. luwenshuni.
一种基于重组尤氏泰勒虫免疫显性蛋白（rTuIP）的酶联免疫吸附测定（ELISA）方法，分别在188份阳性和198份阴性参考血清样本中进行了抗体检测验证。通过双图接收器操作特征（TG-ROC）确定的截断值为32.7%，准确度水平分别为95%和90%。计算得出的诊断敏感性（Se）和特异性（Sp）均为98.4%。对阳性和阴性参考样本的重复性进一步验证表明该检测方法性能可靠。监测实验感染鲁氏泰勒虫的绵羊的抗体动态，结果显示该检测方法能有效检测出感染早期直至感染后两个月针对该病原体的抗体反应。将该检测方法应用于随州和贵阳以前未知的泰勒虫流行地区的现场血清抗体检测，血清阳性率分别为17.8%和11.6%，并且通过在相应血清阳性动物血液中鉴定18S rRNA基因证实了病原体的存在。这些数据支持rTuIP ELISA可能是研究由尤氏泰勒虫和/或鲁氏泰勒虫引起的泰勒虫病流行病学的有用工具。
