【摘要】 通过软件预测猪繁殖与呼吸综合征病毒（PRRSV）N蛋白的主要抗原性表位，利用化学合成的方法合成三条针对N蛋白抗原表位的多肽作为包被抗原，筛选最优的一条建立了检测PRRSV抗体的间接ELISA方法。该ELISA方法的最适条件为：以2.5 μg/mL的合成肽（Pep3）包被酶标板，4℃过夜，不需封闭，阴阳性血清分别稀释200倍，37℃ 作用60 min，辣根过氧化酶（HRP）标记兔抗猪酶标二抗1000倍稀释，37℃作用30 min，TMB 37℃显色15 min，2 mol/L H₂SO₄终止反应，酶标仪A（450 nm）读数。利用所建立的方法测定50份阴性血清的值计算S/P值，求得当S/P值≥0.324时，即为阳性，0.324>样品S/P值>0.212即为可疑，S/P值≤0.212时，即为阴性。之后进行了灵敏度试验，将标准阳性血清进行1∶3200倍稀释仍然呈现阳性，说明该检测方法具有较高的灵敏性；与常见的PCV2、CSFV、PRV、SIV 4种猪疫病阳性血清无交叉反应，说明该检测方法具有较高的特异性；通过对临床的200份样品进行检测，与IDEXX-PRRSV抗体检测试剂盒相比较，符合率达91.4%，说明该校测方法具有较高的符合率。本研究建立的ELISA方法对于PRRSV的常规诊断及流行病学调查具有重要意义。更多还原

【Abstract】 The main antigenic epitope of the N protein of swine reproductive and respiratory syndrome virus （PRRSV） protein was predicted by software analysis,three peptides against the N protein epitope were synthesized as coated antigens,and the best indirect ELISA method for detecting PRRSV antibody was selected.The optimal conditions for the ELISA method is 2.5 μg/mLsynthetic peptide （Pep3） coated with microplate,4℃ overnight,200 times dilution,37℃,horseradish peroxidase （HRP） labeled 1000 times dilution,37℃ for30 min,TMB37℃ color 15 min,2 M H₂SO₄ termination reaction,microplate reader A （450 nm） reading.The S / P value was calculated using the value of 50 negative sera,so that when S / P value of 0.324 is positive,0.324> sample S / P value> 0.212 is suspicious,and S / Pvalue of 0.212 is negative.the sensitivity test was conducted,and the standard positive serum was diluted 1∶3200 times still positive,which indicates the high sensitivity;no cross-reaction with the positive serum of PCV2,CSFV,PRV and SIV,indicating high specificity;through 200 clinical samples,compared with the IDEXX PRRSV antibody detection kit,the compliance rate reached 91.4%,It shows that the calibration method has a high compliance rate.The ELISA method established in this study is important for the routine diagnosis and epidemiological investigation of PRRSV.更多还原