OIE Reference Laboratory for Surra, OIE Collaborating Centre, National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.
Parasitol Res. 2012 Feb;110(2):733-9. doi: 10.1007/s00436-011-2632-9. Epub 2011 Sep 17.
Trypanosoma evansi infection (surra) causes significant losses in livestock production in tropical and sub-tropical areas. The current ELISA recommended by OIE for diagnosis of the disease is based on trypanosome lysate antigen. However, antigenic variation and unstable nature of cell lysate antigen make it difficult to standardize the assay. Thus, there are needs to develop recombinant antigen-based ELISA that improve stability, sensitivity, and specificity of the test. Since tandem repeat (TR) proteins of trypanosomatid parasites generally possess high antigenicity, they have been considered to be the promising antigens for trypanosomosis and leishmaniosis. In this study, IgG responses against 14 recombinant TR proteins of trypanosomes were examined by ELISA. Serum samples were obtained from three water buffaloes experimentally infected with T. evansi. Since Trypanosoma congolense GM6 (TcoGM6) elicited highest IgG responses to all water buffaloes, we further bioinformatically and molecular biologically identified Trypanosoma brucei brucei GM6 (TbbGM6) and T. evansi GM6 (TeGM6) TR genes, respectively. As expected, predicted amino acid sequences of TbbGM6 and TeGM6 were identical while the nucleic acid sequence homology between TbbGM6 and TcoGM6 was 63.8%. All buffaloes became clearly positive in recombinant TbbGM6 (rTbbGM6)-based ELISA at 48 days post-infection, suggesting that rTbbGM6 is usable as a serodiagnostic antigen for chronic T. evansi infection.
伊氏锥虫病(苏拉病)在热带和亚热带地区给畜牧业生产造成了重大损失。目前,OIE 推荐的用于该病诊断的 ELISA 基于锥虫裂解抗原。然而,抗原变异和细胞裂解抗原的不稳定性质使得难以对该检测进行标准化。因此,需要开发基于重组抗原的 ELISA,以提高该检测的稳定性、敏感性和特异性。由于锥虫原生动物的串联重复(TR)蛋白通常具有高抗原性,因此它们被认为是锥虫病和利什曼病的有前途的抗原。在这项研究中,通过 ELISA 检测了针对 14 种锥虫重组 TR 蛋白的 IgG 反应。从三只经实验感染伊氏锥虫的水牛头颈采集血清样本。由于 Trypanosoma congolense GM6(TcoGM6)对所有水牛均引起最高 IgG 反应,因此我们进一步从生物信息学和分子生物学上分别鉴定了 Trypanosoma brucei brucei GM6(TbbGM6)和 T. evansi GM6(TeGM6)的 TR 基因。正如预期的那样,TbbGM6 和 TeGM6 的预测氨基酸序列是相同的,而 TbbGM6 和 TcoGM6 之间的核酸序列同源性为 63.8%。所有水牛在感染后 48 天均在基于重组 TbbGM6(rTbbGM6)的 ELISA 中呈明显阳性,表明 rTbbGM6 可作为慢性 T. evansi 感染的血清学诊断抗原。