Continuous surveillance has shown that H6 subtype avian influenza viruses (AIVs) are prevalent in poultry and occasionally break the species barrier to infect humans. It is therefore necessary to establish a specific, rapid and sensitive method to screen H6 AIVs. In this study, a panel of monoclonal antibodies (mAbs) against the hemagglutinin (HA) of an H6 AIV isolate was produced. The purified mAbs have high affinity and specificity for H6 AIVs. An antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) and immunochromatographic strip were developed based on two mAbs (1D7 and 1F12). The AC-ELISA results showed high sensitivity with a limit of detection (LOD) of 3.9 ng/ml for H6 HA protein and 0.5 HAU (HA units)/100 µl for live H6 subtype AIVs. The average recovery of the AC-ELISA with allantoic fluid, respiratory specimens, and cloacal swabs was 91.907 ± 1.559%, 82.977 ± 1.497% and 73.791 ± 2.588%, respectively. The intra- and inter-assay coefficient of variation was less than 10%. The LOD of immunochromatographic strip was 1 HAU when evaluated by the naked eye, and the detection time was less than 10 min without any equipment. Storage at room temperature or 4 °C for 30 days or 60 days had no effect on sensitivity and specificity of the strip. Thus, the AC-ELISA and immunochromatographic strips described here could be a secondary method to diagnose H6 AIV infections with high specificity, sensitivity, and stability.

中文翻译：

---

基于单克隆抗体检测H6禽流感病毒的抗原捕获酶联免疫吸附测定和免疫色谱带的开发。

连续监测表明，H6亚型禽流感病毒（AIV）在禽类中很普遍，有时会破坏物种屏障以感染人类。因此，有必要建立一种特定，快速和灵敏的方法来筛选H6 AIV。在这项研究中，产生了针对H6 AIV分离株血凝素（HA）的单克隆抗体（mAb）。纯化的mAb对H6 AIV具有高亲和力和特异性。基于两个mAb（1D7和1F12）开发了一种抗原捕获酶联免疫吸附测定（AC-ELISA）和免疫层析带。AC-ELISA结果显示出高灵敏度，H6 HA蛋白的检测限（LOD）为3.9 ng / ml，活H6亚型AIV的检测限为0.5 HAU（HA单位）/ 100 µl。尿囊液，呼吸道标本，泄殖腔拭子分别为91.907±1.559％，82.977±1.497％和73.791±2.588％。批内和批间变异系数小于10％。肉眼评估免疫层析条的检测限为1 HAU，无任何检测时间不到10 min。在室温或4°C下保存30天或60天不会影响试纸条的敏感性和特异性。因此，本文所述的AC-ELISA和免疫色谱条可能是诊断H6 AIV感染的第二种方法，具有很高的特异性，敏感性和稳定性。无需任何设备，检测时间不到10分钟。在室温或4°C下保存30天或60天不会影响试纸条的敏感性和特异性。因此，本文所述的AC-ELISA和免疫色谱条可能是诊断H6 AIV感染的第二种方法，具有很高的特异性，敏感性和稳定性。无需任何设备，检测时间不到10分钟。在室温或4°C下保存30天或60天不会影响试纸条的敏感性和特异性。因此，本文所述的AC-ELISA和免疫色谱条可能是诊断H6 AIV感染的第二种方法，具有很高的特异性，敏感性和稳定性。