| 刊名 | Agricultural Biotechnology |
| 作者 | Zhao LIU1,Bo LIU2, Zhida LIN3, Hang SUN4, Yu SUN4,5*, Xiaohui SONG4* |
| 作者单位 | 1. Dalian Customs, Dalian, Liaoning 116000, China; 2. Changchun Customs, Changchun 130000, China; 3. Gongbei Customs, Zhuhai, Guangdong 519000, China; 4. China Animal Disease Control Center, Beijing 102600, China; 5. Beijing University of Agriculture, Beijing 100096, China |
| DOI | DOI:10.19759/j.cnki.2164-4993.2024.05.007 |
| 年份 | 2024 |
| 刊期 | 5 |
| 页码 | 22-27 |
| 关键词 | Peste des Petits Ruminants; N protein; NH fusion protein; Soluble expression and purification; Rapid quantitative detection |
| 摘要 | [Objectives] This study was conducted to establish a rapid quantitative method for detecting antibody against Peste des Petits Ruminants Virus (PPR V) in sheep serum. [Methods] Soluble N protein and NH fusion protein were obtained in Escherichia coli prokaryotic expression system by optimizing codons and expression conditions of E. coli. Furthermore, based on the purified soluble N protein and NH fusion protein, a high-sensitivity fluorescence immunoassay kit for detecting the antibody against PPR V was established. [Results] The method could quickly and quantitatively detect PPR V antibody in sheep serum, with high sensitivity and specificity, without any cross reaction to other related sheep pathogens. The intra-batch and inter-batch coefficients of variation were less than 10% and 15%, respectively, and the method had good repeatability. Through detection on 292 clinical serum samples, it was compared with the French IDVET competitive ELISA kit, and the coincidence rate of the two methods reached 93.84%. Compared with the serum neutralization test, the detected titer value of the high-sensitivity rapid fluorescence quantitative detection method was basically consistent with the tilter value obtained by the neutralization test on the standard positive serum (provided by the WOAH Brucellosis Reference Laboratory of France). [Conclusions] This method can realize rapid quantitative detection of PPR V antibody on site, and has high practical value and popularization value. |
