Zhang Sujiao, Wang Dongliang, Jiang Yifan, Li Zhoumian, Zou Yawen, Li Meng, Yu Haoyang, Huang Kun, Yang Yi, Wang Naidong
Hunan Provincial Key Laboratory of Protein Engineering in Animal Vaccines, Laboratory of Functional Proteomics, Research Center of Reverse Vaccinology, College of Veterinary Medicine, Hunan Agricultural University, Changsha, 410128, China.
BMC Vet Res. 2019 Mar 6;15(1):79. doi: 10.1186/s12917-019-1810-3.
Porcine circovirus type 3 (PCV3), recently widely isolated from pigs with various clinical conditions, is likely globally epidemic. However, development of serological diagnosis for PCV3 in pigs is ongoing. Our objectives were to: 1) establish an indirect ELISA, using PCV3 capsid protein (Cap) prepared by Baculovirus Expression Vector System (BEVS) as a high-quality coating antigen for detection of PCV3-associated antibodies in serum samples; and 2) use this ELISA to conduct a serological survey for PCV3 in various regions of Hunan province, China.
The PCV3 positive rate to the ELISA assay (total of 190 serum samples) was higher in sows with reproductive failure compared to healthy sows (34/85, 40.0% versus 30/105, 28.6%), with similar results using qPCR assays. Further, in an additional 1038 serum samples collected from January 2016 to May 2018 in various regions of Hunan province and tested with this established ELISA, 20 to 84% were positive for PCV3 (according to region of sera collection), with high PCV3 seroprevalence (> 50%) in herds in Changde, Hengyang and Yueyang. Moreover, among serum samples from herds in Shaoyang and Changde, PCV3 seroprevalence was higher in sows than in other classes of pigs (i.e., suckling piglets, nursery pigs, gilts, growing-finishing pigs and boars).
We developed a full-length PCV3 Cap-based ELISA using a eukaryotic expression system with excellent potential to elucidate PCV3 epidemiology. Based on this assay, PCV3 has been circulating in Hunan province. PCV3 prevalence was lower in healthy sows than in those with reproductive failure. Further studies are warranted to identify the PCV3 responsible for high seroprevalence in sows and determine pathogenesis of PCV3 in sows with reproductive failure.
猪圆环病毒3型(PCV3)最近在患有各种临床病症的猪中广泛分离出来,可能在全球流行。然而,猪PCV3血清学诊断方法仍在开发中。我们的目标是:1)建立一种间接ELISA方法,使用杆状病毒表达载体系统(BEVS)制备的PCV3衣壳蛋白(Cap)作为高质量包被抗原,用于检测血清样本中与PCV3相关的抗体;2)使用该ELISA方法对中国湖南省不同地区的猪进行PCV3血清学调查。
与健康母猪相比,繁殖障碍母猪对ELISA检测(共190份血清样本)的PCV3阳性率更高(34/85,40.0%对30/105,28.6%),qPCR检测结果相似。此外,在2016年1月至2018年5月从湖南省不同地区收集的另外1038份血清样本中,用该建立的ELISA方法检测,20%至84%的样本PCV3呈阳性(根据血清收集地区),常德、衡阳和岳阳的猪群中PCV3血清阳性率较高(>50%)。此外,在邵阳和常德猪群的血清样本中,母猪的PCV3血清阳性率高于其他类别猪(即哺乳仔猪、保育猪、后备母猪、生长育肥猪和公猪)。
我们使用真核表达系统开发了一种基于PCV3全长Cap的ELISA方法,在阐明PCV3流行病学方面具有巨大潜力。基于该检测方法,PCV3已在湖南省流行。健康母猪中PCV3的流行率低于繁殖障碍母猪。有必要进一步研究以确定导致母猪血清阳性率高的PCV3毒株,并确定PCV3在繁殖障碍母猪中的发病机制。