The Development of ELISA for the Detection of the Antibody against Porcine Reproductive and Respiratory Syndrome Virus

• Main Authors: Yi-Ting Huang, 黃儀婷
• Other Authors: 謝明昆
• Format: Others
• Language: zh-TW
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/41075661556725901343

碩士 === 國立中興大學 === 微生物暨公共衛生學研究所 === 103 === Porcine reproductive and respiratory syndrome (PRRS) is an important infectious disease of domestic swine worldwide caused by PRRS virus (PRRSV). The typical clinical signs of PRRS are severe reproductive failure in sows and respiratory problems in pigs of all ages. Currently, ELISA is usually used for the detection of antibody against PRRSV. The advantage of ELISA are time-efficient and cost-effective. The objective of this study was to develop the ELISAs coated with various PRRSV proteins for the detection of antibody against PRRSV. In-house ORF7 ELISA (coated with 15 KDa N protein), Nsp2 ELISA (coated with partial Nsp2 protein), and 14-mer ORF7 ELISA (coated with 14-mer N protein) were used to detect anti-PRRSV antibody. The N protein and Nsp2 protein were expressed by E.coli and purified by affinity chromatography and the 14-mer ORF7 was a 14-mer synthesized peptide. Total 34 pig sera were used for the evaluation and the commercial IDEXX PRRS ELISA was used as gold standard method. The cut-off values were determined by mean plus several standard deviation or the two-graph receiver operation characteristics (TG-ROC) and the TG-ROC gave better results for all three ELISAs. According to the TG-ROC method, the sensitivity, specificity and accuracy were (100 %, 94.10 %, 96.96 %), (91.67 %, 100 %, 96.5 %) and (93.75 %, 88.24 %, 90.9 %) for ORF7 ELISA, Nsp2 ELISA, and 14-mer ORF7 ELISA, respectively. The results showed that ORF7 ELISA had higher sensitivity, specificity and accuracy than those of other in-house ELISAs. ORF7 ELISA is suitable for the detection of anti-PRRSV antibody.