A MONOCLONAL ANTIBODY-BASED ANTIGEN CAPTURE ENZYME-LINKED-IMMUNOSORBENT-ASSAY FOR IDENTIFICATION OF INFECTIOUS-BRONCHITIS VIRUS SEROTYPES

A MONOCLONAL ANTIBODY-BASED ANTIGEN CAPTURE ENZYME-LINKED-IMMUNOSORBENT-ASSAY FOR IDENTIFICATION OF INFECTIOUS-BRONCHITIS VIRUS SEROTYPES
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DOI:
10.1080/03079459308418943
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发表时间:
1993-01-01
期刊:
影响因子:
2.8
通讯作者:
BAUMAN, B
BAUMAN, B
中科院分区:
农林科学3区
文献类型:
--
作者:
NAQI, SA;KARACA, K;BAUMAN, B

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开发了一种抗原捕获酶联免疫吸附测定 (C-ELISA),用于使用对各自血清型的 S1 糖蛋白具有特异性的单克隆抗体 (MAb) 来检测和鉴定阿肯色州、康涅狄格州和马萨诸塞州的传染性支气管炎病毒 (IBV) 血清型。当 S1 特异性 MAb、抗原和鸡血清属于相同血清型时,该测定(设计为双抗体夹心测定)给出最佳结果。然而,当使用群体特异性(M 糖蛋白特异性)MAb 进行抗原捕获时,在抗原和异源鸡血清之间观察到独特的交叉反应模式,表明 IBV M 糖蛋白上表位的复杂分布。仅当使用 M 糖蛋白特异性 MAb 捕获抗原时,用 NP40 处理抗原才能增强 ELISA 信号。尽管 C-ELISA 在检测鸡组织匀浆中的 IBV 方面不一致,但在鸡胚传代一次后,其在检测尿囊液中的病毒方面非常有效。
An antigen-capture enzyme-linked immunosorbent assay (C-ELISA) was developed for detection and identification of infectious bronchitis virus (IBV) serotypes Arkansas, Connecticut, and Massachusetts using monoclonal antibodies (MAbs) specific to the S1 glycoprotein of the respective serotype. The assay (designed as a double-antibody sandwich assay) gave the best results when the S1-specific MAb, antigen, and chicken serum were of the same serotype. However, when a group-specific (M glycoprotein-specific) MAb was used for antigen capture, a distinctive pattern of cross-reactivity was observed between the antigens and heterologous chicken sera, suggesting a complex distribution of epitopes on the IBV M glycoproteins. Treatment of antigen with NP40 enhanced the ELISA signal only when the M glycoprotein-specific MAb was used for antigen capture. Although C-ELISA was inconsistent in detecting IBV in chicken tissue homogenates, it was highly effective in detecting the virus in allantoic fluid after the homogenates were given one chicken embryo passage.