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建立一种竞争化学发光免疫分析方法,使用单克隆抗体识别口蹄疫病毒的 3B 区,用于快速检测口蹄疫病毒感染诱导的抗体。

Development of a competitive chemiluminescence immunoassay using a monoclonal antibody recognizing 3B of foot-and-mouth disease virus for the rapid detection of antibodies induced by FMDV infection.

机构信息

State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China.

出版信息

Virol J. 2021 Sep 26;18(1):193. doi: 10.1186/s12985-021-01663-4.

Abstract

BACKGROUND

Foot-and-mouth disease (FMD) is a devastating animal disease. Anti-non-structural protein (NSP) antibody detection is very important for confirming suspected cases, evaluating the prevalence of infection, certifying animals for trade and controlling the disease.

METHODS

In this study, a competitive chemiluminescence immunoassay (3B-cCLIA) was developed for the rapid detection of antibodies against NSPs in different species of livestock animals using the monoclonal antibody (mAb) 9E2 as a competitive antibody that recognizes NSP 3B.

RESULTS

The cut-off value (50%), diagnostic sensitivity (Dsn) (97.20%, 95.71%, and 96.15%) and diagnostic specificity (Dsp) (99.51%, 99.43%, and 98.36) of the assay were estimated by testing a panel of known-background sera from swine, cattle and sheep, respectively. The accuracy rate of the 3B-cCLIA was further validated and subsequently compared with that of two commercial diagnostic kits. The early diagnostic results showed that antibodies recognizing NSPs developed later (approximately 1-2 days) than antibodies recognizing structural proteins. Furthermore, anti-NSP antibody presence in animals vaccinated multiple times (false positives), especially cattle and sheep, was confirmed, and the false-positive rate increased with the number of vaccinations.

CONCLUSIONS

These results indicate that the 3B-cCLIA is suitable for the rapid detection of antibodies against FMDV NSP 3B in a wide range of species.

摘要

背景

口蹄疫(FMD)是一种毁灭性的动物疾病。抗非结构蛋白(NSP)抗体检测对于确认疑似病例、评估感染流行情况、对动物进行贸易认证和控制疾病非常重要。

方法

本研究使用单克隆抗体(mAb)9E2 作为竞争抗体,针对不同种属的家畜动物中的 NSP 建立了一种快速竞争化学发光免疫分析(3B-cCLIA)。

结果

通过检测一组来自猪、牛和羊的已知背景血清,分别估计了该方法的截断值(50%)、诊断敏感性(Dsn)(97.20%、95.71%和 96.15%)和诊断特异性(Dsp)(99.51%、99.43%和 98.36%)。进一步验证了 3B-cCLIA 的准确性,并随后与两种商业诊断试剂盒进行了比较。早期诊断结果表明,识别 NSP 的抗体出现得较晚(约 1-2 天),而识别结构蛋白的抗体出现得较早。此外,还证实了多次接种(假阳性)的动物存在抗 NSP 抗体,尤其是牛和羊,且随着接种次数的增加,假阳性率也随之增加。

结论

这些结果表明,3B-cCLIA 适用于广泛种属中 FMDV NSP 3B 抗体的快速检测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/be09/8474858/90d1d4751afc/12985_2021_1663_Fig1_HTML.jpg

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