快速诊断家畜日本血吸虫病免疫层析试纸条的研制与初步应用

Development and preliminary application of rapid test strip for domestic animal schistosomiasis japonica

目的开发研制敏感、简便、快速、经济的日本血吸虫病诊断试纸条。方法利用双抗原夹心法,以日本血吸虫可溶性虫卵抗原(SEA)为检测抗原,以胶体金标记SEA为探针,采用自行设计的免疫层析试纸条装置,检测家畜血清中血吸虫特异性IgG抗体,并用ELISA方法进行比较。结果用该试纸条检测107份人工感染血吸虫病羊血清,其检出率为91.6%;检测80份健康绵羊血清,阴性符合率为87.5%;检测20份粪检阳性水牛血清及15份粪检阴性血清,其阳性符合率为100.0%,阴性符合率为86.7%;检测24份肝片形吸虫病羊血清,交叉反应率为12.5%,与锥虫病牛血清未见交叉反应;与ELISA检测结果比较,两种方法具有良好的一致性。结论应用试纸条诊断家畜日本血吸虫病敏感性高、特异性强,且操作简便、快速,不需特殊仪器设备,适合于基层使用。

Objective To develop a simple, rapid, economical and sensitive diagnostic strip for schistosomiasis japonica. Methods The colloidal gold immunochromatography assay （GICA） combined with the double-antigen sandwich assay, the gold immunochromatography assay and colloid gold conjugated antigen technique were used for antibody detection in schistosomiasis japonica. Soluble egg antigen（SEA） of Schistosoma japonicum was conjugated with colloid gold as the detecting probe, and SEA was also coated onto nitrocellulose membrane served as the antigen-antibody complex detecting reagents. Results The strip was used to diagnose schistosomiasis of animals, such as of sheep, buffalo, rat and rabbit. The sensitivity and specificity of this strip were 91.6% and 87.5%. There were 100% positive coincidence rates and 86.7% negative coincidence rates detected 35 sera of buffalo in schistosomaiasis region. It also showed 12.5% cross reactions with sera of schistosomiasis and fascioliasis, and no cross reactions was detected with trypanosomiasis. It showed good coincidence between GICA and ELISA. Conclusions It is evident that this strip （GICA） for detection of specific antibodies against animal schistosomiasis is rapid and simple for handling with high sensitivity and specificity. It is suitable for a wide variety of field applications.