National Centre for Foreign Animal Disease, 1015 Arlington Street, Winnipeg, Manitoba, Canada R3E 3M4.
J Virol Methods. 2009 Dec;162(1-2):194-202. doi: 10.1016/j.jviromet.2009.08.006. Epub 2009 Aug 21.
Nine monoclonal antibodies (mAbs) against avian influenza virus (AI) H5 subtype from mice immunized with inactivated virus H5N1 (A/Turkey/ON/6213/66) were produced. Upon testing, the results indicated that the binding epitopes of eight out of the nine mAbs were conformational, while one mAb (#7) reacted with denatured H5N1 only. Two mAbs #10 and #11 reacted with all of the thirteen H5 strains tested indicating that the binding epitopes of these mAbs were conserved among these H5 subtypes. Possible applications of these mAbs in rapid tests for H5 antigen were explored. Double antibody sandwich (DAS) ELISAs were developed using two selected mAbs #10 and #11. This DAS ELISA detects specific H5 viruses and is able to identify all thirteen H5 strains tested. Three mAbs showed reactivity with AI H5 antigen for both immunofluorescence (IF) and immunohistochemistry. A cELISA used to screen chickens that had been infected with an H5 virus was developed with mAb #9 and recombinant H5 antigen. The sera from chickens that have been infected with an H5N1 virus were examined using the cELISA. 80% of the sera from H5 infected chickens showed a positive H5 specific antibody response at 7 days post-infection (dpi) and remained positive until the end of the experiment on day 30 (>40% inhibition). This panel of the AI H5 specific mAbs is valuable for the development of various immunoassays.
从免疫灭活 H5N1（A/Turkey/ON/6213/66）病毒的小鼠中制备了九种针对禽流感病毒（AI）H5 亚型的单克隆抗体（mAb）。经过测试，结果表明九种 mAb 中有八种的结合表位是构象性的，而一种 mAb（#7）仅与变性的 H5N1 反应。两种 mAb #10 和 #11 与所有测试的 13 种 H5 株均反应，表明这些 mAb 的结合表位在这些 H5 亚型中保守。探讨了这些 mAb 在 H5 抗原快速检测中的可能应用。使用两种选定的 mAb #10 和 #11 开发了双抗体夹心（DAS）ELISA。这种 DAS ELISA 检测特异性 H5 病毒，能够识别所有测试的 13 种 H5 株。三种 mAb 对 AI H5 抗原的免疫荧光（IF）和免疫组织化学均有反应性。使用 mAb #9 和重组 H5 抗原开发了用于筛选感染 H5 病毒的鸡的 cELISA。用 cELISA 检测感染 H5N1 病毒的鸡的血清。感染 H5 的鸡血清在感染后 7 天（dpi）有 80%显示出阳性 H5 特异性抗体反应，并在第 30 天（>40%抑制）实验结束前一直保持阳性。该组 AI H5 特异性 mAb 对于各种免疫测定的开发非常有价值。
