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猪细小病毒(PPV)的重组非结构多聚蛋白NS1作为酶联免疫吸附测定(ELISA)中的诊断抗原,用于区分感染猪和免疫猪。

The recombinant nonstructural polyprotein NS1 of porcine parvovirus (PPV) as diagnostic antigen in ELISA to differentiate infected from vaccinated pigs.

作者信息

Qing L, Lv J, Li H, Tan Y, Hao H, Chen Z, Zhao J, Chen H

机构信息

Laboratory of Animal Virology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, PR China.

出版信息

Vet Res Commun. 2006 Feb;30(2):175-90. doi: 10.1007/s11259-006-3212-9.

Abstract

To differentiate pigs infected with porcine parvovirus (PPV) from those vaccinated with inactivated whole-virus vaccine, an enzyme-linked immunosorbent assay (ELISA) based on detection of a nonstructural polyprotein 1 (NS1) was developed. A threshold of 0.23 optical density units was established and the assay had high specificity (100), sensitivity (88), accuracy (90) and positive predictive value (100) using haemagglutination inhibition as the standard method. A reproducibility test revealed that the coefficients of variation of sera within-plates and between-run were less than 10%. The assay showed no cross-reactivity with antibodies to porcine reproductive respiratory syndrome virus, pseudorabies virus, foot and mouth disease virus, Actinobacillus pleuropneumoniae, Toxoplasma or Chlamydia. Sera obtained from pigs infected with PPV reacted with recombinant NS1 protein in the ELISA. Sera from pigs vaccinated with inactivated whole virus did not recognize this protein in the ELISA. In contrast, antibodies against PPV whole virus were present in both PPV-infected and vaccinated animals. Serum conversion against NS1 was first detected 10 days after infection and NS1-specific antibodies were detectable up to half a year post infection. In conclusion, the PPV-NS1 ELISA can differentiate PPV-infected versus inactivated PPV-vaccinated pigs and could be applied in disease diagnosis and surveillance.

摘要

为了区分感染猪细小病毒(PPV)的猪和接种了灭活全病毒疫苗的猪,开发了一种基于检测非结构多蛋白1(NS1)的酶联免疫吸附测定(ELISA)。以血凝抑制作为标准方法,确定了0.23光密度单位的阈值,该测定具有高特异性(100)、敏感性(88)、准确性(90)和阳性预测值(100)。重复性测试表明,板内和批间血清的变异系数均小于10%。该测定与猪繁殖与呼吸综合征病毒、伪狂犬病病毒、口蹄疫病毒、胸膜肺炎放线杆菌、弓形虫或衣原体的抗体无交叉反应。感染PPV的猪的血清在ELISA中与重组NS1蛋白发生反应。接种灭活全病毒的猪的血清在ELISA中不识别该蛋白。相反,PPV感染和接种疫苗的动物体内均存在抗PPV全病毒的抗体。感染后10天首次检测到针对NS1的血清转化,感染后半年内均可检测到NS1特异性抗体。总之,PPV-NS1 ELISA可以区分感染PPV的猪和接种灭活PPV疫苗的猪,可应用于疾病诊断和监测。

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