重组颗粒抗原GRA1和GRA7用于犬弓形虫感染血清学诊断的评估。
Evaluation of recombinant granule antigens GRA1 and GRA7 for serodiagnosis of Toxoplasma gondii infection in dogs.
作者信息
Wang Zedong, Ge Wei, Huang Si-Yang, Li Jiping, Zhu Xing-Quan, Liu Quan
机构信息
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province 730046, PR China.
出版信息
BMC Vet Res. 2014 Jul 12;10:158. doi: 10.1186/1746-6148-10-158.
BACKGROUND
Toxoplasmosis, caused by the obligate intracellular parasite Toxoplasma gondii, is an important zoonotic disease worldwide. The precise detection of T. gondii infection in dogs has important public health significance. In this study, recombinant granule antigen proteins GRA1 and GRA7 were evaluated as potential diagnostic markers for T. gondii infection in dogs by an indirect enzyme-linked immunosorbent assay (ELISA).
RESULTS
GRA1 and GRA7 were cloned and expressed in Escherichia coli, and the recombinant GRA1, GRA7- and Toxoplasma lysate antigen (TLA)-based ELISAs were developed and evaluated using the canine positive and negative serum samples for anti-T. gondii antibodies determined by modified agglutination test (MAT) and indirect fluorescent antibody test (IFAT), showing a seroprevalence of 15.1% by TLA- and GRA1-ELISA, and 15.8% by GRA7-ELISA, and no significant difference was observed (P > 0.05). When compared with the two reference assays, MAT and IFAT, the GRA7-ELISA showed the highest co-positivity and co-negativity rates. Receiver operating characteristic (ROC) analysis revealed a largest area under curve (AUC) of 0.973 (95% CI, 0.955 to 0.991), and a highest relative sensitivity (93.2%) and specificity (94.0%) for a cut-off value of 0.809 in GRA7-ELISA.
CONCLUSIONS
The results of the present study showed that GRA7-ELISA is highly sensitive and specific, and GRA7 is a potential serodiagnostic marker for the detection of T. gondii infection in dogs.
背景
由专性细胞内寄生虫刚地弓形虫引起的弓形虫病是一种在全球范围内重要的人畜共患病。准确检测犬类中的弓形虫感染具有重要的公共卫生意义。在本研究中，通过间接酶联免疫吸附测定（ELISA）评估重组颗粒抗原蛋白GRA1和GRA7作为犬类弓形虫感染潜在诊断标志物的可能性。
结果
GRA1和GRA7在大肠杆菌中克隆并表达，基于重组GRA1、GRA7和弓形虫裂解物抗原（TLA）开发了ELISA，并使用通过改良凝集试验（MAT）和间接荧光抗体试验（IFAT）确定的犬抗弓形虫抗体阳性和阴性血清样本进行评估，TLA - ELISA和GRA1 - ELISA显示血清阳性率为15.1%，GRA7 - ELISA为15.8%，未观察到显著差异（P>0.05）。与两种参考检测方法MAT和IFAT相比，GRA7 - ELISA显示出最高的共阳性和共阴性率。受试者工作特征（ROC）分析显示，GRA7 - ELISA的曲线下面积（AUC）最大，为0.973（95%CI，0.955至0.991），对于临界值0.809，相对灵敏度最高（93.2%），特异性最高（94.0%）。
结论
本研究结果表明，GRA7 - ELISA具有高度敏感性和特异性，GRA7是检测犬类弓形虫感染的潜在血清学诊断标志物。
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