No more cold-chain failures, using dehydrated reagents in ELISA antibody-detection against animal trypanosomes of African origin
No more cold-chain failures, using dehydrated reagents in ELISA antibody-detection against animal trypanosomes of African origin
复制标题
DOI：
10.1016/j.vetpar.2021.109568
复制
发表时间：
2021-09-06
影响因子：
2.6
通讯作者：
Desquesnes, M.
中科院分区：
文献类型：
--
作者：
Bossard, G.;Millogo, L.;Desquesnes, M.
非洲锥虫引起的动物锥虫病（ATAO）主要由刚果锥虫萨凡纳型（TCS）、T. brucei brucei（TBB）、T.间日疟原虫（TV）和T.伊文思病是影响家养和野生哺乳动物的广泛疾病，并具有巨大的经济影响。ATAO临床怀疑通常通过寄生虫学和分子学方法确认，而血清流行病学调查通常使用OIE推荐的基于纯化锥虫全细胞溶解物可溶性抗原（WCLSA）的ELISA方法进行;该试剂通常冷冻储存。为了扩大这种ELISA检测，我们评估，标准化，并验证使用脱水而不是冷冻WCLSA和血清样本。对于三种ELISA测定（TV、TCS和TBB），重复性研究显示重复之间没有显著差异。使用冷冻而非冻干抗原和血清获得的结果与Pearson相关值（>0.93）和Lin测量值（“非常好”至“极好”）强相关。重现性是稳健的，技术员间效应的Pearson相关值>0.97，实验室间测试的Pearson相关值为0.87（TV）至0.97（TBB和TCS）;根据Lin的测量，它们的组合为“非常满意”至“优秀”，对定性测试结果没有影响。脱水试剂的优点是可在室温下运输，从而能够安全地向区域实验室提供试剂。脱水试剂与ATAO（/OIE）标准诊断方案纲要一起，将使流行国家能够在区域一级对ATAO进行血清学诊断。非洲国家最近推出的锥虫渐进控制途径中的这一非常受欢迎的改进可能会在不久的将来扩展到拉丁美洲。
Animal trypanosomoses due to trypanosomes of African origin (ATAO), mainly caused by Trypanosoma congolense type Savannah (TCS), T. brucei brucei (TBB), T. vivax (TV), and T. evansi, are widespread diseases that affect domestic and wild mammals and have a huge economic impact. ATAO clinical suspicions are usually confirmed by parasitological and molecular methods, while sero-epidemiological surveys are generally carried out using the OIE-recommended ELISA method based on whole cell lysate soluble antigens (WCLSA) from purified trypanosomes; this reagent is usually stored frozen. With a view to expanding this ELISA test, we assessed, standardized, and validated the use of dehydrated rather than frozen WCLSA and serum samples. For the three ELISA assays (TV, TCS & TBB), a repeatability study revealed no significant difference between repeats. The results obtained using frozen rather than freeze-dried antigen and serum strongly correlated for Pearson's correlation values (>0.93) and Lin's measure ("very good" to "excellent"). Reproducibility was robust, with Pearson's correlation values >0.97 for inter technician effects, and 0.87 (TV) to 0.97 (TBB & TCS) for inter-laboratory tests; their combination was "very satisfactory" to "excellent" according to Lin's measure and there was no impact on qualitative test results. Dehydrated reagents offer the advantage of shipment at room temperature, allowing the secured provision of reagents to regional laboratories. Together with a compendium of standard diagnostic protocols for ATAO (/OIE), dehydrated reagents will enable the serological diagnosis of ATAO at regional level in endemic countries. This very welcome improvement in the context of the Progressive Control Pathway for trypanosomes, recently launched by African countries, will possibly be extended to Latin America in the near future.