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本研究建立了一种用PVC薄膜作载体、将前鞭毛体可溶性抗原包被在PVC凹板上,采用HRP-SPA作第二抗体及无毒的TMB作底物的快速ELISA方法,该法简便、经济、实用。与经典ELISA平行对照检测24份骨髓穿刺证实的内脏利什曼病犬血清,结果:快速ELISA法阳性符合率为100％(24/24),而经典ELISA法为95.83％(23/24);对47份采自非流行区的正常犬血清,两法均仅有1例呈假阳性反应,占2.13％;用快速ELISA法检测实验感染了旋毛虫、肺吸虫、钩虫、弓形虫等犬血清共26份,均未见交叉反应。用该法又调查了四川汶川县黑热病重流行区的124份犬血清,同时进行骨髓涂片检查,结果快速ELISA查出阳性犬39只,占31.45％(39/124),骨髓涂片查见利什曼原虫者有24只,占19.35％(24/124),这24只犬血清对快速ELISA法均呈阳性反应。
In this study, we established a rapid ELISA method using PVC membrane as carrier, coating promastigote soluble antigen on PVC concave plate, using HRP-SPA as secondary antibody and nontoxic TMB as substrate, which is simple and convenient ,Economical and practical. In parallel with classical ELISA, the serum of 24 patients with visceral leishmaniasis confirmed by bone marrow aspiration was detected. Results: The positive coincidence rate of fast ELISA was 100% (24/24), while that of classical ELISA was 95.83% (23/24) 47 normal dogs sera collected from non-endemic areas, only 1 case of two methods were false positives, accounting for 2.13%; using rapid ELISA assay experiment infected with Trichinella, paragonimiasis, hookworm, toxoplasma canine serum 26, no cross-reaction. Using this method, we investigated 124 canine serums in the meningitis area of Wenchuan County, Sichuan Province. Meanwhile, bone marrow smears were also performed. The results showed that 39 dogs (31.45%) were positive by rapid ELISA (39/124) Leishmania there are 24, accounting for 19.35% (24/124), these 24 canine serum for the rapid ELISA were positive.