肺吸虫抗体快速检测试剂盒(金标渗滤法)的研制和应用

Development of rapid diagnostic Kit(Dot Immunogold Filtration Assay) for detection of antibodies angainst Paragonimus westermani

目的开发研制简易、快速、经济、准确的肺吸虫病诊断试剂盒。方法以肺吸虫成虫可溶性蛋白为检测用抗原,以胶体金标记羊抗人IgG为探针,采用自行设计的渗滤装置,检测病人血清中肺吸虫特异性IgG抗体;用深圳康百得生物技术有限公司提供的肺吸虫抗体ELISA检测试剂盒作比较。结果用本试剂盒检测90人份肺吸虫病人血清和100人份健康人血清,其敏感性为98.9%(89/90),特异性为100%(100/100),Youden’s指数为0.989。用该试剂盒分别检测25人份血吸虫病和华支睾吸虫病患者血清,交叉反应率为24%(6/25)和4%(1/25);与姜片虫、囊虫、丝虫、肺结核病人血清均未见交叉反应。ELISA检测结果比较,符合率为93.5%,无显著性差异(χ2=0.7949,P>0.05)。结论肺吸虫抗体快速检测试剂盒(金标渗滤法)敏感性高、特异性强,可与ELISA相媲美,且操作更为简便、快速,结果判读容易,不需特殊仪器设备,非常适合临床检验和现场查病。

To devdope a simple, rapid, economical and sensitive diagnostic kit for paragonimiasis, crude extracts of adult worms of Paragonimus westermani were used as antigen and goat anti-human IgG antibodies were conjugated with colloid gold as the detecting probe. A special pad for dot immunogold filtration assay （DIGFA） was designed according to the reaction procedure. An paragonimiasis-ELISA kit was utilized to detect the antibodies in the paraUel sera with this rapid kit. The sensitivity and specificity of this kit were 98.9 % （89/90） and 100 % （100/100） respectively. There were 24 % （6/25） and 4 % （1/25） cross reactions with sera of shistosomiasis and clonorchlasis. No cross reactions was detected with tuberculosis and other helminthiasis, such as fasciolopsiais, cysticercosis, filariasis. There was no significant difference in comparison with the detecting results by using ELISA kit （x^2 = 0.7949, P 〉0.05）. It is evident this diagnostic kit （DIGFA） for detection of specific antibodies against Paragonimus westermani is rapid and simple for handling with high sensitivity and specificity. That is very useful to clinical diagnosis and field investigation of paragonimia- sis.