Evaluation of an epitope-blocking enzyme-linked immunosorbent assay for the detection of antibodies to influenza A virus in domestic and wild avian and mammalian species
Evaluation of an epitope-blocking enzyme-linked immunosorbent assay for the detection of antibodies to influenza A virus in domestic and wild avian and mammalian species
复制标题
DOI：
10.1016/j.jviromet.2009.06.001
复制
发表时间：
2009-10-01
影响因子：
3.1
通讯作者：
Root, J. Jeffrey
中科院分区：
文献类型：
--
作者：
Sullivan, Heather J.;Blitvich, Bradley J.;Root, J. Jeffrey
开发了一种表位阻断酶联免疫吸附试验（bELISA），用于检测分类学上不同的家养和野生脊椎动物物种中的甲型流感病毒抗体。与之前发表的需要试剂生产、最终用户操作或尚未评价用于哺乳动物和禽类物种的bELISA不同，本试验使用市售重组核蛋白抗原和相应的核蛋白特异性单克隆抗体进行，并已显示对多种禽类和哺乳动物物种有效。使用从实验感染的绿头鸭（Anas platyrhynchos）和浣熊（Procyon lotor）获得的251个血清样品，将bELISA作为血清筛选测定的功效与琼脂凝胶免疫扩散（AGID）测定进行比较。AGID法与bELISA法的一致性为94.1%（95%CI = 89.9。98.3），浣熊和71.2%（95% CI = 63.5，78.9）的野鸭和82.8%（95% CI = 78.2，87.3）的整体。在实验性感染研究的早期时间点和较高的系列稀释度下检测甲型流感病毒抗体，证明bELISA比AGID测定法更灵敏。还使用来自6个禽类物种和6个哺乳动物物种的另外745份血清样本，将bELISA监测天然甲型流感病毒暴露的有效性与AGID测定进行了比较。这种bELISA为大规模监测分类学上不同的脊椎动物物种中的甲型流感病毒暴露提供了一种快速、可靠和廉价的技术。出版社：Elsevier B.V.
An epitope-blocking enzyme-linked immunosorbent assay (bELISA) was developed for the detection of antibodies to influenza A virus in taxonomically diverse domestic and wild vertebrate species. In contrast to the bELISAs published previously that require reagent production, manipulation by the end-user, or have not been evaluated for use with both mammalian and avian species, this assay is performed using commercially available recombinant nucleoprotein antigen and corresponding nucleoprotein-specific monoclonal antibody and has been shown to work with multiple avian and mammalian species. The efficacy of the bELISA as a serum screening assay was compared to the agar gel immunodiffusion (AGID) assay using 251 serum samples obtained from experimentally infected mallards (Anas platyrhynchos) and raccoons (Procyon lotor). The concordance between the AGID assay and bELISA was 94.1% (95% Cl = 89.9. 98.3) for raccoons, and 71.2% (95% Cl = 63.5, 78.9) for mallards and 82.8% (95% Cl = 78.2, 87.3) overall. The bELISA was more sensitive than the AGID assay as demonstrated by the detection of antibodies to influenza A virus at earlier time points in experimental infection studies and at higher serial dilutions. The efficacy of the bELISA to monitor natural influenza A virus exposure was also compared to the AGID assay using an additional 745 serum samples from six avian species and six mammalian species. This bELISA provides a rapid, reliable, and inexpensive technique for large-scale surveillance of influenza A virus exposure in taxonomically diverse vertebrate species. Published by Elsevier B.V.