›› 2018, Vol. 45 ›› Issue (4): 881-887.doi: 10.16431/j.cnki.1671-7236.2018.04.006

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Establishment and Application of Sandwich ELISA for Quantitative Detection of Porcine Reproductive and Respiratory Syndrome Virus Soluble Receptors

XIA Wenlong1, WU Zhi2, GUO Changming2, ZHU Shanyuan2, YU Shupei1, ZHANG Xinyu1, XIA Xiaoli1, SUN Huaichang1   

  1. 1. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China;
    2. Jiangsu Key Laboratory for High-tech Research and Development of Veterinary Biopharmaceuticals, Jiangsu Animal Husbandry and Veterinary College, Taizhou 225300, China
  • Revised:2017-12-01 Online:2018-04-20 Published:2018-04-25


To establish a sandwich ELISA for quantitative detection of porcine reproductive and respiratory syndrome virus (PRRSV) soluble receptors,porcine sialoadhesin (Sn) soluble receptor Sn4D-Fc was purified from the recombinant adenovirus-transduced PK-15 cell and used as the standard antigen,mouse anti-porcine Sn serum was used as the first antibody and biotin-labeled rabbit anti-porcine Sn polyclonal antibody as the second antibody.The established sandwich ELISA was validated by detection of the soluble receptor in the serum samples of rAd-Sn4D-Fc-injected pigs.The results showed that the Sn4D-Fc standard antigen was purified to a purity of 94.3%,and was able to be recognized by anti-Sn serum.By using 5 μg/mL of the first antibody and 1:4 000 of second antibody,the detection limit of established sandwich ELISA was up to 0.4 ng/mL without cross reaction to control antigen.The Sn4D-Fc soluble receptor could be detected in the rAd-injected pigs with the highest expression level of 6.54 ng/mL and the longest duration of 15 d.These data suggested that the established sandwich ELISA was usable for quantitative detection of PRRSV soluble receptors in vitro and in vivo.

Key words: porcine reproductive and respiratory syndrome virus (PRRSV); soluble receptor; sandwich ELISA; quantitative detection

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