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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Development of an Indirect Tams1 Enzyme-Linked Immunosorbent Assay for Diagnosis of Theileria annulata Infection in Cattle
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Development of an Indirect Tams1 Enzyme-Linked Immunosorbent Assay for Diagnosis of Theileria annulata Infection in Cattle

机译：间接Tams1酶联免疫吸附测定技术在牛环蒂勒虫感染诊断中的应用

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摘要

An enzyme-linked immunosorbent assay (ELISA) was developed based on a recombinant major Theileria annulata merozoite surface antigen, Tams1. Four different recombinant proteins derived from two different Tams1 alleles, both in two different truncated forms, were tested for their performance in the ELISA. Furthermore, antigen concentration, various buffers, washing protocol, and the choice of anti-total-immunoglobulin G (IgG), anti-IgG1, or anti-IgG2 as second antibody were evaluated. The performance of the resulting ELISA was analyzed by measuring the coefficient of variation (CV). A total of 22 sera were analyzed over the measurement range, resulting in a CV of ca. 10%, whereas 30% variation is the maximum acceptable. The cutoff value was determined by the two-graph receiver operating characteristic (TG-ROC), using the indirect fluorescent antibody test (IFAT) as a reference. It was shown that up to 3 months postinfection (p.i.) IFAT is more sensitive and specific, whereas beyond 3 months p.i. ELISA performed as well as IFAT. The cutoff was determined at maximal sensitivity, based on the TG-ROC after 3 months p.i. Nine calves experimentally infected with four different T. annulatastocks remained positive in the ELISA for at least 1 year p.i. Finally, limited cross-reaction was found only with T. parvaantisera, but not with any other Theileria orBabesia species. Since the T. parva endemic area hardly overlaps with T. annulata, the Tams1 ELISA has the potential to become a useful tool in the epidemiology of tropical theileriosis.
机译：基于重组主要的 Theileria annulata 裂殖子表面抗原Tams1，开发了一种酶联免疫吸附试验（ELISA）。在ELISA中测试了来自两种不同截短形式的两种不同Tams1等位基因的四种不同重组蛋白的性能。此外，还评估了抗原浓度，各种缓冲液，洗涤方案以及抗-总免疫球蛋白G（IgG），抗-IgG1或抗-IgG2作为第二抗体的选择。通过测量变异系数（CV）分析所得ELISA的性能。在整个测量范围内对总共22个血清进行了分析，得出CV为。 10％，而30％的变化是最大可接受值。使用间接荧光抗体测试（IFAT）作为参考，通过两幅图接收器操作特性（TG-ROC）确定临界值。结果表明，感染后3个月内（p.i.）IFAT更为敏感和特异，而感染后3个月以上。 ELISA和IFAT一样执行。于3个月p.i后根据TG-ROC以最高灵敏度确定临界值。九头牛实验性地感染了四种不同的 T。环孢菌种在ELISA中至少每年保持阳性1年。最终，仅与 T发现了有限的交叉反应。 parva antisera，但不能与其他 Theileria 或 Babesia 物种一起使用。自 T。 parva 流行区几乎不与 T重叠。环线虫，Tams1 ELISA有望成为热带回旋虫病流行病学中有用的工具。

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