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Establishment and Preliminary Application of ELISA Method of PRRSV Antibody by Using Synthetic Peptides as Antigen

SUN Jian;Weihai Ocean Vocational College;Institute of Special Economic Animal and Plant Sciences,CAAS;  
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 H_2SO_4 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.
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