4 Verification of post-chemotherapeutic clearance of Theileria equi through concordance of nested PCR and immunoblot
4 Verification of post-chemotherapeutic clearance of Theileria equi through concordance of nested PCR and immunoblot
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DOI：
10.1016/j.ttbdis.2017.08.007
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发表时间：
2018-02-01
影响因子：
3.2
通讯作者：
Knowles, D. P.
中科院分区：
文献类型：
--
作者：
Wise, L. N.;Kappmeyer, L. S.;Knowles, D. P.
通过进口限制和监测，包括美国在内的某些国家仍然没有马焦虫病等特定传染病的流行。流行地区通常采用预免疫作为控制疾病的主要方法，但在非流行国家，需要化学绝育结合确认寄生虫消除的方法来维持无病状态。二丙酸亚胺威 (ID) 能够清除受感染马匹的持续性马泰勒虫感染，这一能力已通过接受治疗的马匹无法通过血液转移传播而得到证实。然而，抗 T 蛋白普遍存在较长时间的持续存在。 Equi 抗体可使 cELISA 或 IFA 等监管测试长时间保持阳性。持续的阳性检测给兽医监管和国际贸易带来了挑战。使用针对 ema1 基因的巢式聚合酶链式反应 (nPCR) 与测量抗 EMA1 和抗 EMA2 抗体下降的免疫印迹 (IB) 之间的一致性来验证从 179 匹 ID 治疗的马中清除马锥虫。这些数据支持使用 IB 来证明在成功进行 ID 治疗后，马 T. 感染的马中抗 EMA1 和 EMA2 滴度下降。此类数据为阴性 nPCR 提供了一致的支持，并允许更及时地确定马锥虫的有效 ID 清除。 ID 治疗后结果表明，虽然 nPCR 在 14 天时始终呈阴性，cELISA 在 1 年后通常仍呈阳性，但免疫印迹在 4 个月后平均呈阴性，与 nPCR 100% 一致。
Certain countries including the United States remain non-endemic for particular infectious diseases such as equine piroplasmosis through import restrictions and surveillance. Endemic regions often employ premunition as the primary method to control disease, however in non-endemic countries, chemosterilization combined with methods to confirm parasite elimination are required to maintain disease-free status. The ability of imidocarb diproprionate (ID) to clear persistent Theileria equi infection from infected horses has been shown through the inability of treated horses to transmit via blood transfer. However, the common lengthy persistence of anti-T. equi antibody causes regulatory tests such as cELISA or IFA to remain positive for extended periods. Persistence of positive testing creates challenges for regulatory veterinary medicine and international trade. Concordance between nested polymerase chain reaction (nPCR) targeting the ema1 gene and immunoblotting (IB) measuring declination in anti-EMA1 and anti-EMA2 antibody were used to verify clearance of T. equi from 179 ID-treated horses. These data support the use of IB to demonstrate declining anti-EMA1 and EMA2 titers in T. equi-infected horses subsequent to successful ID treatment. Such data provide concordant support to a negative nPCR and allow for a more timely determination of effective ID clearance of T. equi. The post ID treatment results indicate that while nPCR was consistently negative by 14 days and cELISA generally remained positive after 1 year, immunoblot was on average negative after 4 months and 100% in agreement with nPCR.