Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
Instituto de Farmacologia e Biologia Molecular, Universidade Federal de São Paulo, São Paulo, São Paulo, Brazil.
Vet Parasitol. 2021 Aug;296:109495. doi: 10.1016/j.vetpar.2021.109495. Epub 2021 Jun 15.
Human trypanosomiases and animal trypanosomoses are caused by distinct protozoan parasites of the genus Trypanosoma. The etiological agents of bovine trypanosomosis (BT) are T. vivax, T. congolense, or T. brucei, whose acute infections are initially characterized by hyperthermia, following moderate to severe anemia, subcutaneous edema, lethargy, reduced milk production, progressive weight loss, enlarged lymph nodes, reproductive disorders and death. Animals that survive the acute phase might recover and progress to the chronic, often asymptomatic, phase of infection. Despite their low sensitivity due to the characteristic low parasitemia, simple and costless direct parasitological examinations are the preferred diagnostic methods for animals. Thus, most of the epidemiological studies of BT are based on serological techniques using crude antigen. In this study, we describe the use of the MyxoTLm recombinant protein as an antigen on serological assays. Anti-T. vivax IgM and anti-T. vivax IgG ELISA assays using purified MyxoTLm revealed specificity rates of 91.30 % and 95.65 % and sensitivity rates of 82.35 % and 88.23 %, respectively, being higher than reported for crude antigens. Also, MyxoTLm demonstrated a good performance to detect IgM (ROC curve area = 0.8568) and excellent performance to detect IgG (ROC curve area = 0.9565) when compared to a crude antigen. T. evansi crude antigen used in the indirect anti-T. vivax IgM ELISA reached 70.58 % sensitivity and 78.26 % specificity, and had a lower test performance (ROC curve area = 0.7363). When applied to the anti-T. vivax IgG ELISA, the crude antigen reached 82.35 % sensitivity and 69.56 % specificity, also presenting a low performance with area under the ROC curve of 0.7570. Therefore, the use of MyxoTLm as an antigen on serological diagnosis of BT revealed to increase the sensitivity and the specificity if compared to crude antigens.
人体锥虫病和动物锥虫病是由不同的原生动物寄生虫引起的。牛锥虫病(BT)的病原体是 T. vivax、T. congolense 或 T. brucei,其急性感染最初表现为高热,随后出现中度至重度贫血、皮下水肿、昏睡、产奶量减少、体重逐渐减轻、淋巴结肿大、生殖障碍和死亡。在急性阶段幸存下来的动物可能会康复并进入慢性、通常无症状的感染阶段。尽管由于特征性的低寄生虫血症导致其敏感性较低,但简单且廉价的直接寄生虫学检查仍然是动物的首选诊断方法。因此,大多数 BT 的流行病学研究都是基于使用粗抗原的血清学技术。在这项研究中,我们描述了使用 MyxoTLm 重组蛋白作为血清学检测中的抗原。使用纯化的 MyxoTLm 进行抗 T. vivax IgM 和抗 T. vivax IgG ELISA 检测,其特异性分别为 91.30%和 95.65%,敏感性分别为 82.35%和 88.23%,高于粗抗原报道的结果。此外,与粗抗原相比,MyxoTLm 在检测 IgM(ROC 曲线面积为 0.8568)和 IgG(ROC 曲线面积为 0.9565)时表现出良好的性能。用于间接抗 T. vivax IgM ELISA 的 T. evansi 粗抗原的敏感性为 70.58%,特异性为 78.26%,检测性能较低(ROC 曲线面积为 0.7363)。当应用于抗 T. vivax IgG ELISA 时,粗抗原的敏感性为 82.35%,特异性为 69.56%,ROC 曲线下面积也较低,为 0.7570。因此,与粗抗原相比,将 MyxoTLm 作为 BT 血清学诊断中的抗原使用,可以提高敏感性和特异性。