Development of a Novel Double Antibody Sandwich Quantitative Enzyme-Linked Immunosorbent Assay for Detection of Porcine Epidemic Diarrhea Virus Antigen.
Development of a Novel Double Antibody Sandwich Quantitative Enzyme-Linked Immunosorbent Assay for Detection of Porcine Epidemic Diarrhea Virus Antigen.
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新型双抗体夹心定量酶联免疫吸附法检测猪流行性腹泻病毒抗原的开发
DOI：
10.3389/fvets.2020.540248
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发表时间：
2020
影响因子：
3.2
通讯作者：
Li B
中科院分区：
文献类型：
--
作者：
Fan B;Sun J;Zhu L;Zhou J;Zhao Y;Yu Z;Sun B;Guo R;He K;Li B
猪流行性腹泻病毒（PEDV）可引起哺乳仔猪急性腹泻和脱水，死亡率高。本研究利用抗PEDV N蛋白的单克隆抗体和抗PEDV兔血清，建立了检测PEDV的双抗体夹心定量酶联免疫吸附试验（DAS-qELISA）。用DAS-qELISA检测PEDV N蛋白的最低检测限为1 μg/L，病毒滴度为102.0 TCID 50/ml。然后使用DAS-qELISA和逆转录酶PCR（RT-PCR）筛选总共90个肠样品和237个粪便样品中PEDV的存在。DAS-qELISA的特异性为98.1%，敏感性为93.5%。DAS-qELISA与RT-PCR的准确率为95.7%。更重要的是，通过使用DAS-qELISA，在一次灭活疫苗制备前后，病毒抗原浓度保持不变。结果表明，DAS-qELISA可用于灭活疫苗样品和临床样品的抗原检测。为PEDV疫苗的评价和疾病诊断提供了新的方法。
Porcine epidemic diarrhea virus (PEDV) causes acute diarrhea and dehydration in sucking piglets with a high mortality rate. Here, we developed a double antibody sandwich quantitative enzyme-linked immunosorbent assay (DAS-qELISA) for detection of PEDV using a specific monoclonal antibody against PEDV N protein and anti-PEDV rabbit serum. Using DAS-qELISA, the detection limit of recombinant PEDV N protein and virus titer were approximately 1 μg/L and 102.0 TCID50/ml, respectively. A total of 90 intestinal and 237 fecal samples were then screened for the presence of PEDV using DAS-qELISA and reverse transcriptase PCR (RT-PCR). DAS-qELISA had a high specificity of 98.1% and sensitivity of 93.5%. The accuracy rate between DAS-qELISA and RT-PCR was 95.7%. More importantly, the viral antigen concentrations remained unchanged before and after one inactivated vaccine preparation by using the DAS-qELISA. These results suggest DAS-qELISA could be used for antigen detection of inactivated vaccine samples and clinical samples. It is a novel method for diagnosing diseases and evaluation of the PEDV vaccine.
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DOI：
10.1089/mab.2013.0023
发表时间：
2013-10-01
影响因子：
--
作者：
Li, Bin;Du, Luping;He, Kongwang
通讯作者：
He, Kongwang
影响因子：
4.8
作者：
Pan Y;Tian X;Li W;Zhou Q;Wang D;Bi Y;Chen F;Song Y
通讯作者：
Song Y
影响因子：
6.4
作者：
Huang YW;Dickerman AW;Piñeyro P;Li L;Fang L;Kiehne R;Opriessnig T;Meng XJ
通讯作者：
Meng XJ
影响因子：
3.1
作者：
Kim YK;Lim SI;Cho IS;Cheong KM;Lee EJ;Lee SO;Kim JB;Kim JH;Jeong DS;An BH;An DJ
通讯作者：
An DJ
影响因子：
2.4
作者：
Sozzi E;Luppi A;Lelli D;Martin AM;Canelli E;Brocchi E;Lavazza A;Cordioli P
通讯作者：
Cordioli P