Foot-and-Mouth Disease Virus Antigen Detection Enzyme-Linked Immunosorbent Assay Using Multiserotype-Reactive Monoclonal Antibodies
Foot-and-Mouth Disease Virus Antigen Detection Enzyme-Linked Immunosorbent Assay Using Multiserotype-Reactive Monoclonal Antibodies
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DOI：
10.1128/jcm.00695-09
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发表时间：
2009-11-01
影响因子：
9.4
通讯作者：
Sakamoto, Kenichi
中科院分区：
文献类型：
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作者：
Morioka, Kazuki;Fukai, Katsuhiko;Sakamoto, Kenichi
开发了基于单克隆抗体（MAb）的夹心直接酶联免疫吸附试验（MSD-ELISA）方法，可以检测口蹄疫病毒（FMDV）抗原，包括多血清型（MSD-ELISA/MS）（针对O、A、C和Asia 1）和单血清型（MSD-ELISA/SS）（具体针对O、A和Asia 1）。MAb 1H 5用作抗原捕获抗体，其与FMDV的所有七种血清型反应。在MSD-ELISA/MS和MSD-ELISA/SS中，MAb 71 F2、70 C4、16 C6和7 C2分别用作多血清型（O、A、C和Asia 1）、O型、A型和Asia 1型的过氧化物酶标记检测抗体。我们的MSD-ELISA显示出高度的特异性。他们产生了非常低背景的阴性样品（缓冲液、血浆和唾液），并且能够从临床样品（血浆和唾液）中检测FMDV抗原，结果与实时逆转录PCR的结果相关。在灵敏度方面，MSD-ELISA显示出比间接夹心ELISA方法更高的针对每种稀释血清型抗原的光密度值，间接夹心ELISA方法目前在世界动物卫生组织手册中推荐。MSD-ELISA的敏感性和特异性足以用于口蹄疫病毒的抗原诊断。
Monoclonal antibody (MAb)-based sandwich direct enzyme-linked immunosorbent assay (MSD-ELISA) methods that can detect foot-and-mouth disease virus (FMDV) antigens, both multiserotype (MSD-ELISA/MS) (for O, A, C, and Asia 1) and single-serotype (MSD-ELISA/SS) (for O, A, and Asia 1, specifically), were developed. MAb 1H5 was used as an antigen-trapping antibody that reacted with all seven serotypes of FMDV. The MAbs 71F2, 70C4, 16C6, and 7C2 were used as peroxidase-labeled detecting antibodies for multiple serotypes (O, A, C, and Asia 1), type O, type A, and type Asia 1, respectively, in both MSD-ELISA/MS and MSD-ELISA/SS. Our MSD-ELISAs showed high specificity. They produced a very low background of negative samples (buffer, plasma, and saliva) and were able to detect FMDV antigens from clinical samples (plasma and saliva), with results correlating with those of real-time reverse transcription-PCR. In terms of sensitivity, the MSD-ELISAs showed higher optical density values against each diluted serotype antigen than the indirect sandwich ELISA method, which is currently recommended in the manual of the World Organization for Animal Health. The sensitivity and specificity of the MSD-ELISAs seem to be sufficient for the antigenic diagnosis of FMDV.