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合成肽酶免疫测定法在区分反刍动物呼吸道合胞病毒两个亚组中的验证

Validation of synthetic peptide enzyme immunoassays in differentiating two subgroups of ruminant respiratory syncytial virus.

作者信息

Grubbs S T, Kania S A, Potgieter L N

机构信息

Department of Comparative Medicine, College of Veterinary Medicine, University of Tennessee, Knoxville 37901-1071, USA.

出版信息

J Vet Diagn Invest. 2001 Mar;13(2):123-7. doi: 10.1177/104063870101300205.

Abstract

Subgroup-specific peptide-based enzyme immunoassays from each respective G-glycoprotein of the ovine and the bovine respiratory syncytial virus (RSV) were developed to detect RSV-specific IgG responses in cattle. Antigenic peptides from the respective G-glycoprotein were identified from the extracellular central hydrophobic region (amino acids 158-189) located between 2 mucin-rich regions. These antigenic peptides identified by epitope mapping from each G-glycoprotein were synthesized and used to develop the subgroup-specific enzyme immunoassays. The negative cutoff for each enzyme immunoassay was established as the mean optical density of indirect immunofluorescent antibody-negative bovine sera plus 3 SDs. The sensitivity (82.9%) and specificity (100%) of the bovine enzyme immunoassay and the specificity (95.8%) of the ovine enzyme immunoassay were determined by comparison with indirect immunofluorescence (used as the "gold standard"). The negative and positive predictive values were calculated for each assay. The presence of serum antibody to ovine RSV in cattle implies that this virus infects cattle and may contribute to the pathogenesis of bovine respiratory disease.

摘要

我们开发了基于绵羊和牛呼吸道合胞病毒(RSV)各自G糖蛋白的亚组特异性肽酶免疫测定法,以检测牛的RSV特异性IgG反应。从位于两个富含粘蛋白区域之间的细胞外中央疏水区域(氨基酸158 - 189)中鉴定出各自G糖蛋白的抗原肽。通过对每个G糖蛋白进行表位作图鉴定出的这些抗原肽被合成,并用于开发亚组特异性酶免疫测定法。每种酶免疫测定法的阴性临界值确定为间接免疫荧光抗体阴性牛血清的平均光密度加上3个标准差。通过与间接免疫荧光法(用作“金标准”)比较,确定了牛酶免疫测定法的灵敏度(82.9%)和特异性(100%)以及绵羊酶免疫测定法的特异性(95.8%)。计算了每种测定法的阴性和阳性预测值。牛体内存在抗绵羊RSV的血清抗体表明该病毒可感染牛,并可能导致牛呼吸道疾病的发病机制。

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