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一种用于检测口蹄疫病毒 A 型的简单快速胶体金免疫层析试纸条检测方法。

A simple and rapid colloidal gold-based immunochromatogarpic strip test for detection of FMDV serotype A.

机构信息

Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China.

出版信息

Virol Sin. 2011 Feb;26(1):30-9. doi: 10.1007/s12250-011-3166-5. Epub 2011 Feb 18.

Abstract

A sandwich format immunochromatographic assay for detecting foot-and-mouth disease virus (FMDV) serotypes was developed. In this rapid test, affinity purified polyclonal antibodies from Guinea pigs which were immunized with sucking-mouse adapted FMD virus (A/AV88(L) strain) were conjugated to colloidal gold beads and used as the capture antibody, and affinity purified polyclonal antibodies from rabbits which were immunized with cell-culture adapted FMD virus (A/CHA/09 strain) were used as detector antibody. On the nitrocellulose membrane of the immunochromatographic strip, the capture antibody was laid on a sample pad, the detector antibody was printed at the test line(T) and goat anti-guinea pigs IgG antibodies were immobilized to the control line(C). The lower detection limit of the test for a FMDV 146S antigen is 11.7 ng/ml as determined in serial tests after the strip device was assembled and the assay condition optimization. No cross reactions were found with FMDV serotype C, Swine vesicular disease (SVD), Vesicular stomatitis virus (VSV) and vesicular exanthema of swine virus (VES) viral antigens with this rapid test. Clinically, the diagnostic sensitivity of this test for FMDV serotypes A was 88.7% which is as same as an indirect-sandwich ELISA. The specificity of this strip test was 98.2% and is comparable to the 98.7% obtained with indirect-sandwich ELISA. This rapid strip test is simple, easy and fast for clinical testing on field sites; no special instruments and skills are required, and the result can be obtained within 15 min. To our knowledge, this is the first rapid immunochromatogarpic assay for serotype A of FMDV.

摘要

建立了一种用于检测口蹄疫病毒(FMDV)血清型的夹心免疫层析分析方法。在这个快速检测中,用经适应鼠适应的 FMD 病毒(A/AV88(L)株)免疫的豚鼠亲和纯化的多克隆抗体被偶联到胶体金颗粒上作为捕获抗体,用适应细胞培养的 FMD 病毒(A/CHA/09 株)免疫的兔亲和纯化的多克隆抗体作为检测抗体。在免疫层析条的硝酸纤维素膜上,将捕获抗体铺在样品垫上,将检测抗体打印在测试线上(T),并将山羊抗豚鼠 IgG 抗体固定在对照线上(C)。在组装条装置并优化检测条件后,通过系列检测确定该检测方法对 FMDV 146S 抗原的最低检测限为 11.7 ng/ml。该快速检测与 FMDV 血清型 C、猪水疱病(SVD)、水疱性口炎病毒(VSV)和猪水疱病病毒(VES)病毒抗原无交叉反应。临床研究表明,该检测方法对 FMDV 血清型 A 的诊断灵敏度为 88.7%,与间接夹心 ELISA 相同。该条带试验的特异性为 98.2%,与间接夹心 ELISA 获得的 98.7%相当。该快速条带检测方法简单、易于现场临床检测,不需要特殊仪器和技能,结果可在 15 分钟内获得。据我们所知,这是第一个用于检测 FMDV 血清型 A 的快速免疫层析分析方法。

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