Grego E, Uslenghi F, Strasser M, Luzzago C, Frigerio M, Peletto S, Rosati S
Dipartimento di Produzioni Animali, Epidemiolgia ed Ecologia, Facoltà di Medicina Veterinaria, Via Leonardo da Vinci 44, 10095 Grugliasco (TO), Italy.
J Vet Diagn Invest. 2007 Jan;19(1):21-7. doi: 10.1177/104063870701900104.
The BVDV envelope glycoprotein E(rns)/gp48 and the C terminal 79 amino acids of the capsid protein coding region were expressed in a baculovirus system and antigenically characterized. Western blot assay was used to detect recombinant E(rns) (r-E(rns)) in infected insect cells using specific monoclonal antibodies. The r-E(rns) was then used in an indirect ELISA to detect BVDV specific antibodies in a panel of 540 well-characterized sera. Results of the r-E(rns) ELISA were compared to those obtained with a commercially available competitive ELISA targeting anti-NS2/3 antibodies. A good correlation was observed between the 2 ELISA (kappa = 0.916, 95% C.I.: 0.876, 0.956). Using the commercial NS2/3 ELISA as the reference test, the relative sensitivity of r-E(rns) ELISA was 97.5% (95% C.I.: 94.3%, 99.1%) and the relative specificity was 93.9% (95% C.I.: 89.4%, 96.9%), while relative specificity was 100% (95% C.I.: 97%, 100%) using true negative sera (derived from a negative herd). All but 1 antigen positive animals (n = 36) tested negative in the r-E(rns) ELISA; among them all 22 confirmed PI animals were negative by r-E(rns) ELISA. The ability of r-E(rns) ELISA to identify cattle immunized with inactivated vaccine was also demonstrated in a small group of cattle, compared to an NS2/3 antibody ELISA. Results suggest that r-E(rns) ELISA represents an alternative test for antibody generated by natural infection or BVDV vaccination.
牛病毒性腹泻病毒（BVDV）包膜糖蛋白E(rns)/gp48及衣壳蛋白编码区的C末端79个氨基酸在杆状病毒系统中表达并进行了抗原特性分析。使用特异性单克隆抗体，通过蛋白质免疫印迹法检测感染昆虫细胞中的重组E(rns)（r-E(rns)）。然后将r-E(rns)用于间接酶联免疫吸附测定（ELISA），以检测540份特征明确的血清中的BVDV特异性抗体。将r-E(rns) ELISA的结果与使用针对抗NS2/3抗体的市售竞争性ELISA获得的结果进行比较。两种ELISA之间观察到良好的相关性（kappa = 0.916，95%置信区间：0.876，0.956）。以市售NS2/3 ELISA作为参考检测方法，r-E(rns) ELISA的相对敏感性为97.5%（95%置信区间：94.3%，99.1%），相对特异性为93.9%（95%置信区间：89.4%，96.9%），而使用真阴性血清（来自阴性牛群）时相对特异性为100%（95%置信区间：97%，100%）。除1只抗原阳性动物（n = 36）外，所有动物在r-E(rns) ELISA中检测均为阴性；其中所有22只确诊的持续性感染（PI）动物在r-E(rns) ELISA中均为阴性。与NS2/3抗体ELISA相比，在一小群牛中也证明了r-E(rns) ELISA识别用灭活疫苗免疫的牛的能力。结果表明，r-E(rns) ELISA是一种用于检测自然感染或BVDV疫苗接种产生的抗体的替代检测方法。
