摘要
为评估猪群血清中猪繁殖与呼吸综合征病毒(PRRSV)中和抗体水平,本研究以原核表达的重组HP-PRRSV GP5蛋白为包被抗原,通过对反应条件进行优化,建立了检测GP5抗体的间接ELISA(r GP5-ELISA)。结果显示该方法与猪圆环病毒、猪瘟病毒、猪伪狂犬病毒、细小病毒、日本乙型脑炎病毒阳性血清均无交叉反应,具有良好的特异性。重复性试验结果显示,组内组间变异系数均小于5.3%。应用建立的r GP5-ELISA方法和IDEXX-ELISA对288份临床血清样品进行检测,结果显示两种方法具有较低的相关性(R=0.831)。中和试验(SN)结果显示,r GP5-ELISA检出阳性血清的中和效价显著高于IDEXX-ELISA(p<0.01)。Western blot复检r GP5-ELISA检测呈阴性的65份猪血清样品,其中58份呈阴性、7份为阳性;而IDEXX-ELISA检测则该65份血清全部呈阳性。对比SN和western blot试验,r GP5-ELISA比商品化IDEXX-ELISA具有更高的SN抗体符合率,因而其检测结果能够更好地反映猪群血清中和抗体水平。本研究建立的方法可以用于大规模猪群PRRSV中和抗体水平评估,并为PRRS流行病学调查提供了技术支持。
To simplify the evaluation of neutralizing antibody level against porcine reproductive and respiratory syndrome virus （PRRSV） in porcine sera from pig herds, an indirect rGP5-ELISA was developed with recombinant GP5 of highly pathogenic PRRSV （HP-PRRSV） as coating antigen. The detection results indicated that the rGP5-ELISA was specific to PRRSV and had a Rood repeatability （CV〈5.30%）. A total of 288 serum samples from pigs were comparatively detected by the established rGP5-ELISA and IDEXX-ELISA, a commercial detection kit for antibody to PRRSV, and paralleled by serum neutralization （SN） test and western blot （WB） to analyze correlation between the two methods and SN test. Then, the assay results revealed that the rGP5-ELISA was specific for PRRSV antiserum detection, but a low correlation was showed between S/P values of the two methods （R=0.831）. In addition, the SN assay displayed that the rGP5-ELISA detected a significant higher titer of PRRSV-specific neutralizing antibodies than that of IDEXX-ELISA （p〈0.01）. Furthermore, the 65 serum negative samples detected by rGPS-ELISA were verified by WB, of which only 7 samples were positive; while, all of the 65 samples were positive by IDEXX-ELISA verification. Obviously, the results of rGPS-ELISA detections could better reflectthe actual antibody level against PRRSV in pigs. Therefore, the rGPS-ELISA has the potential to be used for developing a reliable kit to detect PRRSV-specific antibodies and simplify the evaluation of the neutralizing ability of the corresponding antiserum induced by PRRSV infection or vaccine.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2015年第10期774-779,共6页
Chinese Journal of Preventive Veterinary Medicine
基金
国家自然科学基金(31170146)
