猪带绦虫六钩蚴TSOL18重组蛋白快速诊断试纸条研制

目的以猪带绦虫六钩蚴重组蛋白(TSOL18)为抗原,建立一种简便快速的猪囊虫抗体检测方法,评价其血清学诊断的价值。方法用胶体金颗粒标记纯化的TSOL18重组蛋白,将兔抗TSOL18-GST-IgG和TSOL18重组抗原分别喷涂于硝酸纤维素膜的质控线和检测线上,与PVC垫板等部件按顺序装配成猪囊虫抗体快速检测试纸条;用该试纸条检测猪血清样品测定敏感性和特异性。结果该试纸条与全囊虫抗原ELISA的相对敏感性和特异性分别为75.0%(12/16)和95.2%(40/42),与剖检计数法的相对敏感性达80.0%(12/15)。试纸条在4℃存放12个月,检测结果稳定,重复性好。结论基于TSOL18建立的胶体金免疫层析试纸条操作简单、快速敏感、稳定性好,可用于猪囊虫病感染的诊断和筛查。

Development of gold immunochromatographic strip using the recombinant protein TSOL18 for rapid diagnosis of swine cysticercosis

The objective of the present study was to establish a simple and rapid method to detect swine cysticercosis antibodies by using the 18 kDa recombinant antigen TSOL18 of Taenia solium oncosphere as antigen in simple colloid gold immunochromatographic assay （GICA） for the rapid diagnosis of swine eysticercosis and to evaluate its serodiagnostie value. Colloid gold coupled with recombinant purified TSOL18 protein was dispensed on the glass fiber to prepare the gold-labled TSOL18 protein and was coated on nitrocellulose membrane as the capture line. The rabbit anti= TSOL18--GST IgG was used as the control line. This rapid gold immunochromatographic strip was assembled in regular sequence through the different accessories being sticked on PVC board. Compared with EI,ISA using crude cyst fluid antigens, the relative sensitivity and specificity of the strip was 75.0 %（12/16） and 95.2%（40/42）, respectively. Its relative sensitivity compared with the parasitologieal examination during necropsy was 80.0 % （12/15） , and its stability and reproducibility were quite excellent after storage of the strip at 4℃ for 12 months. It is evident that the gold immunochromatographis strip using the recombinant TSOL18 protein appears to be a simple, rapid and sensitive method for the rapid diagnosis and screening of swine cysticercosis.