Latha D, Geetha M, Ramadass P, Narayanan R B
Centre for Biotechnology, Anna University, Guindy, Chennai 600025, India.
Vet Immunol Immunopathol. 2007 Oct 15;119(3-4):278-86. doi: 10.1016/j.vetimm.2007.06.006. Epub 2007 Jun 17.
An IgM-ELISA based on a 16-kDa recombinant protein produced for the conserved and functional middle region of nucleocapsid protein of Canine distemper virus was developed. Out of 70 serum samples from distemper-suspected and vaccinated dogs analyzed, 34 serum samples (49%) were positive. The specificity of this ELISA was confirmed by blocking and adsorption experiments. The IgM-ELISA based on the recombinant nucleocapsid protein showed a strong correlation (r=0.857, p<0.0001 at 95% CI) and good agreement (kappa=0.714) with the conventional Vero cell culture distemper antigen based IgM-ELISA. The percent positivity was more in dogs with systemic signs (62%) by recombinant nucleocapsid protein IgM-ELISA. Out of 70 clinical serum samples, 69 samples were used along with 4 control sera used in the IgM-ELISA for the detection of viral RNA by Slot blot hybridization and 26 of them (36%) were positive. Fifty-one percent agreement was observed between the recombinant nucleocapsid protein IgM-ELISA and Slot blot hybridization. The analysis of clinical history of the dogs with systemic signs supported the application of IgM-ELISA over Slot blot hybridization in the early detection of distemper infection.
基于犬瘟热病毒核衣壳蛋白保守且具有功能的中间区域产生的16 kDa重组蛋白开发了一种IgM - ELISA。在分析的70份来自疑似犬瘟热和接种过疫苗的犬的血清样本中，34份血清样本（49%）呈阳性。通过阻断和吸附实验证实了该ELISA的特异性。基于重组核衣壳蛋白的IgM - ELISA与基于传统Vero细胞培养犬瘟热抗原的IgM - ELISA显示出强相关性（r = 0.857，95%置信区间下p < 0.0001）和良好的一致性（kappa = 0.714）。通过重组核衣壳蛋白IgM - ELISA检测，有全身症状的犬的阳性率更高（62%）。在70份临床血清样本中，69份样本与IgM - ELISA中使用的4份对照血清一起用于通过狭缝印迹杂交检测病毒RNA，其中26份（36%）呈阳性。重组核衣壳蛋白IgM - ELISA与狭缝印迹杂交之间的一致性为51%。对有全身症状的犬的临床病史分析支持在犬瘟热感染的早期检测中，IgM - ELISA比狭缝印迹杂交更适用。
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