Langedijk J P, Middel W G, Schaaper W M, Meloen R H, Kramps J A, Brandenburg A H, van Oirschot J T
Department of Bovine Virology, Institute for Animal Science and Health (ID-DLO), Lelystad, Netherlands.
J Immunol Methods. 1996 Jun 21;193(2):157-66. doi: 10.1016/0022-1759(96)00039-7.
Peptides deduced from the central hydrophobic region (residues 158-189) of the G protein of bovine and ovine respiratory syncytial virus (RSV) and of human RSV subtypes A and B were synthesized. These peptides were used to develop ELISAs to measure specifically antibodies against these types and subtypes of RSV. We have evaluated the bovine RSV-G peptide in both an indirect ELISA and in a blocking ELISA. Specificity and sensitivity, relative to a routine diagnostic ELISA that detects antibodies against the RSV F-protein in bovine sera, were 98% and 92% respectively for the indirect peptide-based ELISA, and 98% and 98% for the blocking peptide-based ELISA. In paired serum samples, rises in antibody titer were detected more frequently with the indirect peptide-based ELISA than with the routine F-ELISA. Furthermore, the peptide-based G-ELISAs were able to differentiate between antibodies against BRSV and HRSV, and between those against BRSV and ORSV. In addition, the indirect peptide-based ELISA was selective for HRSV subtype A and B antibodies. This study shows that peptides, corresponding to the central hydrophobic region of the attachment protein G of several RSVs, can be used successfully as antigens in highly specific and sensitive immunoassays.
合成了从牛和羊呼吸道合胞病毒(RSV)以及人RSV A和B亚型的G蛋白中央疏水区域(第158 - 189位氨基酸残基)推导而来的肽段。这些肽段被用于开发酶联免疫吸附测定(ELISA),以特异性检测针对这些RSV类型和亚型的抗体。我们在间接ELISA和阻断ELISA中对牛RSV - G肽进行了评估。相对于检测牛血清中针对RSV F蛋白抗体的常规诊断ELISA,基于肽段的间接ELISA的特异性和灵敏度分别为98%和92%,基于肽段的阻断ELISA的特异性和灵敏度分别为98%和98%。在配对血清样本中,基于肽段的间接ELISA比常规F - ELISA更频繁地检测到抗体滴度升高。此外,基于肽段的G - ELISA能够区分针对牛RSV(BRSV)和人RSV(HRSV)的抗体,以及针对BRSV和羊RSV(ORSV)的抗体。另外,基于肽段的间接ELISA对HRSV A和B亚型抗体具有选择性。本研究表明,与几种RSV的附着蛋白G的中央疏水区域相对应的肽段可成功用作高度特异性和灵敏免疫测定中的抗原。