快速检测 H7 亚型禽流感病毒的免疫层析条的研制。
Development of an immunochromatographic strip for rapid detection of H7 subtype avian influenza viruses.
机构信息
College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450002, China.
Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou, 450002, China.
出版信息
Virol J. 2021 Apr 7;18(1):68. doi: 10.1186/s12985-021-01537-9.
BACKGROUND
H7N9 avian influenza virus (AIV) including highly and low pathogenic viruses have been detected in China since 2013. H7N9 AIV has a high mortality rate after infection in humans, and most human cases have close contacted with poultry in the live poultry market. Therefore, it is necessary to develop a rapid point-of-care testing (POCT) technique for H7N9 AIV detection.
METHODS
The H7N9 AIV was inactivated and purified, and was used as the antigen to immunize BALB/c. Twelve H7-HA specific monoclonal antibodies (McAbs) were produced through the hybridoma technique. The McAb 10A8 was conjugated with colloid gold as detecting antibody; McAb 9B6 was dispensed on the nitrocellulose membran as the capture test line and the Goat-anti mouse IgG antibody was dispensed as control line respectively. The immunochromatographic strip was prepared.
RESULTS
The analysis of ELISA and virus neutralization test showed that the obtained McAbs specifically recognized H7 HA. Based on the prepared strip, the detection of H7 AIV was achieved within 10 min. No cross-reaction occurred between H7 AIVs and other tested viruses. The detection limit of the strip for H7 was 2.4 logEID/0.1 mL for chicken swab samples.
CONCLUSION
The McAbs were specific for H7 and the immunochromatographic strip developed in this study was convenient, rapid and reliable for the detection of H7 AIV. The strip could provide an effective method for the rapid and early detection of H7 AIV.
背景
自 2013 年以来，中国已检测到 H7N9 禽流感病毒（AIV），包括高致病性和低致病性病毒。H7N9 AIV 感染人类后的死亡率很高，大多数人类病例都与活禽市场中的家禽有密切接触。因此，有必要开发一种用于 H7N9 AIV 检测的快速即时检测（POCT）技术。
方法
将 H7N9 AIV 灭活并纯化，用作抗原免疫 BALB/c。通过杂交瘤技术生产了 12 株 H7-HA 特异性单克隆抗体（McAb）。将 McAb 10A8 与胶体金缀合作为检测抗体；将 McAb 9B6 分配在硝酸纤维素膜上作为捕获测试线，山羊抗小鼠 IgG 抗体作为对照线。制备免疫层析条。
结果
ELISA 和病毒中和试验分析表明，获得的 McAb 特异性识别 H7 HA。基于制备的条带，在 10 分钟内即可实现 H7 AIV 的检测。H7 AIV 与其他测试病毒之间没有交叉反应。该条带对鸡拭子样本中 H7 的检测限为 2.4 logEID/0.1 mL。
结论
这些 McAb 特异性识别 H7，本研究开发的免疫层析条方便、快速、可靠，可用于检测 H7 AIV。该条带可提供一种快速早期检测 H7 AIV 的有效方法。
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