Kashyap S Phani, Hiremath Jagadish, Vinutha S, Patil Sharanagouda S, Suresh Kuralayanapalya P, Roy Parimal, Hemadri Divakar
ICAR-National Institute of Veterinary Epidemiology and Disease Informatics, P.B. No. 6450, Yelahanka, Bengaluru, Karnataka, India.
Department of Microbiology and Biotechnology, Jain University, Bengaluru, Karnataka, India.
Vet World. 2020 Dec;13(12):2587-2595. doi: 10.14202/vetworld.2020.2587-2595. Epub 2020 Dec 5.
Porcine reproductive and respiratory syndrome (PRRS) is a disease endemic in many countries and is of economic importance. India was free from PRRS until the first outbreak was reported from a North-East Indian state in 2013. Since then, disease outbreaks have been reported from North-East India and the pilot study conducted earlier showed that it is gradually spreading to the rest of India. Considering there are no locally developed population screening tests available for PRRS and imported diagnostic/screening tests are expensive, the present study was aimed at developing recombinant nucleocapsid (rN) protein-based indirect enzyme-linked immunosorbent assay (iELISA).
The rN protein of PRRS virus (PRRSV) was produced following standard cloning, expression, and purification procedures. Using this antigen, iELISA was optimized for the detection of serum antibodies to PRRSV. The sensitivity and specificity of the test were assessed by comparing it with a commercial PRRSV antibody detection kit.
A total of 745 serum samples from ten different states of India were screened using the developed iELISA. The iELISA had a relative specificity of 76.18% and sensitivity of 82.61% compared to the commercial ELISA (Priocheck PRRSV ELISA kit, Thermo Fisher Scientific, USA).
The iELISA, which deployed rN protein from Indian PRRSV, was found to be suitable in the serological survey and may be a useful tool in future disease surveillance programs.
猪繁殖与呼吸综合征（PRRS）在许多国家呈地方流行性，具有重要经济意义。印度在2013年东北部一个邦报告首例疫情之前一直没有PRRS。自那时起，印度东北部报告了疫情，早期开展的初步研究表明，该病正在逐渐蔓延至印度其他地区。鉴于目前没有印度本土开发的用于PRRS的群体筛查检测方法，且进口诊断/筛查检测方法昂贵，本研究旨在开发基于重组核衣壳（rN）蛋白的间接酶联免疫吸附测定（iELISA）。
按照标准的克隆、表达和纯化程序制备PRRS病毒（PRRSV）的rN蛋白。使用该抗原，对iELISA进行优化以检测针对PRRSV的血清抗体。通过与商用PRRSV抗体检测试剂盒比较评估该检测方法的敏感性和特异性。
使用所开发的iELISA对来自印度十个不同邦的745份血清样本进行了筛查。与商用ELISA（美国赛默飞世尔科技公司的Priocheck PRRSV ELISA试剂盒）相比，该iELISA的相对特异性为76.18%，敏感性为82.61%。
发现采用来自印度PRRSV的rN蛋白的iELISA适用于血清学调查，可能是未来疾病监测计划中的一种有用工具。
