Development and validation of an ELISA kit (YF MAC-HD) to detect IgM to yellow fever virus
Development and validation of an ELISA kit (YF MAC-HD) to detect IgM to yellow fever virus
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DOI：
10.1016/j.jviromet.2015.08.025
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发表时间：
2015-12-01
影响因子：
3.1
通讯作者：
Johnson, Barbara W.
中科院分区：
文献类型：
--
作者：
Basile, Alison Jane;Goodman, Christin;Johnson, Barbara W.
黄热病病毒（YFV）是世界热带和亚热带地区的地方病，非洲每年约有18万人感染。血清学检测是确定暴发和通知开始接种运动的决定的主要实验室诊断手段。世界卫生组织向非洲参考实验室分发了YFV检测试剂，美国疾病控制和预防中心负责生产和提供这些试剂。CDC M抗体捕获ELISA是一种2天检测，在收到新批次时需要滴定试剂，这导致检测不一致和材料浪费。在这里，我们描述了基于CDC方法的试剂盒检测（YF MAC-HD）的开发，该方法在约3.5小时内完成，可疑样品被重新分配至过夜方案。与2天测试相比，该试剂盒显示>90%的准确性。该套件设计用于最少的设备，并储存在4摄氏度，消除了对冷冻能力的需要。该试剂盒能够耐受暂时的次优储存条件，这将缓解运输或断电问题，并且证明了>6个月的保质期，准确性没有恶化。运行YF MAC-HD所需的所有试剂均包含在试剂盒中，并且是一次性使用的，每个试剂盒有8或24个样品选项。设想在不久的将来进行实地试验，这将使该方法得到改进。使用YF MAC-HD有望减少材料浪费，提高流行区YFV血清学检测的质量和一致性。由爱思唯尔公司出版
Yellow fever virus (YFV) is endemic in tropical and sub-tropical regions of the world, with around 180,000 human infections a year occurring in Africa. Serologic testing is the chief laboratory diagnostic means of identifying an outbreak and to inform the decision to commence a vaccination campaign. The World Health Organization disseminates the reagents for YFV testing to African reference laboratories, and the US Centers for Disease Control and Prevention (CDC) is charged with producing and providing these reagents. The CDC M-antibody capture ELISA is a 2-day test, requiring titration of reagents when new lots are received, which leads to inconsistency in testing and wastage of material. Here we describe the development of a kit-based assay (YF MAC-HD) based upon the CDC method, that is completed in approximately 3.5 h, with equivocal samples being reflexed to an overnight protocol. The kit exhibits >90% accuracy when compared to the 2-day test. The kits were designed for use with a minimum of equipment and are stored at 4 degrees C, removing the need for freezing capacity. This kit is capable of tolerating temporary sub-optimal storage conditions which will ease shipping or power outage concerns, and a shelf life of >6 months was demonstrated with no deterioration in accuracy. All reagents necessary to run the YF MAC-HD are included in the kit and are single-use, with 8 or 24 sample options per kit. Field trials are envisioned for the near future, which will enable refinement of the method. The use of the YF MAC-HD is anticipated to reduce materials wastage, and improve the quality and consistency of YFV serologic testing in endemic areas. Published by Elsevier B.V.