Jain Beenu, Lambe Upendra, Tewari Anuj, Kadian Surender Kumar, Prasad Minakshi
1Department of Veterinary Microbiology, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana 125004 India.
2Department of Animal Biotechnology, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana 125004 India.
Virusdisease. 2018 Jun;29(2):192-198. doi: 10.1007/s13337-018-0450-8. Epub 2018 Apr 17.
Foot and mouth disease is an economically important transboundary disease of wildlife and cloven hoofed animals including ruminants. In the absence of vaccination, detection of antibodies against structural proteins (SPs) of foot-and-mouth disease virus is an indicator of infection. In the present study, a rapid dot blot assay using gold nanoparticlees was developed for the detection of antibodies against SPs of FMDV. Commercially available FMD vaccine was used as a source of FMD antigen. After the synthesis of gold nanoparticles (GNPs), the GNP-dot blot assay was tested and was found very sensitive, as the detection of antibody was up to 10 of serum dilution. The GNP-dot assay was found specific as it didn't give dot with normal horse sera, fetal bovine sera and neonatal bovine calf serum samples when tested at 10 working dilution. When 30 serum samples from post-vaccinated buffaloes were tested at dilution of 10, all the samples were found positive with the variable intensity of dot. The synthesized GNPs and conjugated GNPS with antibody were characterized for their absorption limit, for their stability and for their approximate size. These characterized conjugated and non-conjugated GNPs were also analyzed by Transmission electron microscopy and Scanning electron microscopy. The GNP dot blot assay developed in this work gave promising results using vaccine antigen and can form an important tool for rapid diagnosis of FMD in FMD free countries, zones free of FMD and during last stage of FMD eradication when FMD vaccination will be withdrawn.
口蹄疫是一种对野生动物和包括反刍动物在内的偶蹄动物具有重要经济影响的跨界疾病。在没有接种疫苗的情况下,检测针对口蹄疫病毒结构蛋白(SPs)的抗体是感染的一个指标。在本研究中,开发了一种使用金纳米颗粒的快速斑点印迹法来检测针对口蹄疫病毒结构蛋白的抗体。市售的口蹄疫疫苗用作口蹄疫抗原的来源。在合成金纳米颗粒(GNPs)后,对口蹄疫病毒结构蛋白金纳米颗粒斑点印迹法进行了测试,发现其非常灵敏,抗体检测可达血清稀释度为1:10。口蹄疫病毒结构蛋白金纳米颗粒斑点印迹法具有特异性,在1:10工作稀释度下测试时,正常马血清、胎牛血清和新生牛犊血清样本均未出现斑点。当对30份接种疫苗后的水牛血清样本进行1:10稀释度测试时,所有样本均呈阳性,斑点强度各不相同。对合成的金纳米颗粒以及与抗体偶联的金纳米颗粒进行了吸收极限、稳定性和近似尺寸的表征。还通过透射电子显微镜和扫描电子显微镜对这些表征的偶联和未偶联金纳米颗粒进行了分析。本研究中开发的口蹄疫病毒结构蛋白金纳米颗粒斑点印迹法使用疫苗抗原取得了有前景的结果,可成为在无口蹄疫国家、口蹄疫无疫区以及在口蹄疫根除的最后阶段(届时将停止口蹄疫疫苗接种)快速诊断口蹄疫的重要工具。