定量检测猪繁殖与呼吸综合征病毒游离受体夹心ELISA方法的建立与应用

doi:10.16431/j.cnki.1671-7236.2018.04.006

Abstract

Abstract:

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

CLC Number:

S852.61⁺4

XIA Wenlong, WU Zhi, GUO Changming, ZHU Shanyuan, YU Shupei, ZHANG Xinyu, XIA Xiaoli, SUN Huaichang. Establishment and Application of Sandwich ELISA for Quantitative Detection of Porcine Reproductive and Respiratory Syndrome Virus Soluble Receptors[J]. , 2018, 45(4): 881-887.

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

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