Senthilkumaran Chandrika, Yang Ming, Bittner Hilary, Ambagala Aruna, Lung Oliver, Zimmerman Jeffrey, Giménez-Lirola Luis G, Nfon Charles
Canadian Food Inspection Agency, National Centre for Foreign Animal Disease (NCFAD), Winnipeg, Manitoba (Senthilkumaran, Yang, Bittner, Ambagala, Lung, Nfon); College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA (Zimmerman, Giménez-Lirola); Department of Animal Science, University of Manitoba, Winnipeg, Manitoba (Nfon).
Can J Vet Res. 2017 Apr;81(2):82-90.
Virus nucleic acids and antibody response to pathogens can be measured using swine oral fluids (OFs). Detection of foot-and-mouth disease virus (FMDV) genome in swine OFs has previously been demonstrated. Virus isolation and viral antigen detection are additional confirmatory assays for diagnosing FMDV, but these methods have not been evaluated using swine OF. The objectives of this study were to further validate the molecular detection of FMDV in oral fluids, evaluate antigen detection and FMDV isolation from swine OFs, and develop an assay for isotypic anti-FMDV antibody detection in OFs. Ribonucleic acid (RNA) from FMDV was detected in OFs from experimentally infected pigs by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) from 1 day post-infection (dpi) to 21 dpi. Foot-and-mouth disease virus (FMDV) was isolated from OFs at 1 to 5 dpi. Additionally, FMDV antigens were detected in OFs from 1 to 6 dpi using a lateral flow immunochromatographic strip test (LFIST), which is a rapid pen-side test, and from 2 to 3 dpi using a double-antibody sandwich enzyme-linked immunosorbent assay (DAS ELISA). Furthermore, FMDV-specific immunoglobulin A (IgA) was detected in OFs using an isotype-specific indirect ELISA starting at dpi 14. These results further demonstrated the potential use of oral fluids for detecting FMDV genome, live virus, and viral antigens, as well as for quantifying mucosal IgA antibody response.
可使用猪口腔液(OFs)来检测病毒核酸和对病原体的抗体反应。此前已证实可在猪口腔液中检测口蹄疫病毒(FMDV)基因组。病毒分离和病毒抗原检测是诊断FMDV的其他确证性检测方法,但这些方法尚未在猪口腔液中进行评估。本研究的目的是进一步验证口腔液中FMDV的分子检测,评估从猪口腔液中进行抗原检测和FMDV分离,并开发一种用于检测口腔液中同型抗FMDV抗体的检测方法。通过定量实时逆转录聚合酶链反应(qRT-PCR),在感染后1天(dpi)至21 dpi期间,从实验感染猪的口腔液中检测到了FMDV的核糖核酸(RNA)。在1至5 dpi时从口腔液中分离出了口蹄疫病毒(FMDV)。此外,使用侧向流动免疫层析试纸条检测(LFIST,一种快速的床边检测方法)在感染后1至6 dpi时检测到了口腔液中的FMDV抗原,使用双抗体夹心酶联免疫吸附测定(DAS ELISA)在感染后2至3 dpi时检测到了该抗原。此外,从感染后第14天开始,使用同型特异性间接ELISA在口腔液中检测到了FMDV特异性免疫球蛋白A(IgA)。这些结果进一步证明了口腔液在检测FMDV基因组、活病毒和病毒抗原以及定量黏膜IgA抗体反应方面的潜在用途。