抗杜氏利什曼原虫犬分离株单克隆抗体的研制及用于病犬血清循环抗原的检测

Preparation of Monoclonal Antibodies against Leishmania Donovani Canine Isolate and Detection of Circulating Antigens in Sera of Dogs with Leishmaniasis Based on McAbs

本文报道用杜氏利什曼原虫大分离株前鞭毛体膜抗原免疫BALB/c小鼠的脾细胞与Sp2/0瘤细胞融合,得到多株分泌抗体较高且稳定的细胞株。进一步筛选出McAb C11-G10-A4可用于检测病犬血清循环抗原。用McAb-AST检测采自甘肃省黑热病流行区的30份犬血清,阳性率30％。与骨髓涂片查病原体的总符合率为86.7％,阳性符合率达100％。69份非流行区对照大血清检测结果,仅1份出现阳性反应,可见本试验的敏感性较高,特异性较强(98.55％)。

BALB/c mice were immunized with the membrane antigen produced from Leishmania donovani promastigotes and its spleen cells were fused with the nonproducer mouse myeloma Sp2/0 cells to generate hybridomas screcting McAbs against L. donovani. Our results showed that the hybridoma growth efficiency for the fusion was 98.9%, of which 58.1% of the hybridomas specifically secreted antibodies against L. donovani. Three hybridomas secreting antibodies highly and stably were obtained through twice cloning. Using McAb C11-G10-A4, we detected the sera of 30 dogs lived in endemic districts in Wen County, Gansu province by McAb-AST, of which 0 were positive with 30% positive rate, furthermore, each of the dogs was examined microscopically using the bone marrow smear method to determine if there L. donovani parasites existed. Comparison of the positive rate between the two methods showed that the coincidental rate between them reached 86.7%. of control sera from 69 dogs lived in non-endemic districts only 1 case was found to be positive. The results showed that the specificity of McAb-AST is high, up to 98.55%.