Rajamanickam Hema Sayee, Madhusudan Hosamani , Narayanan Krishnaswamy, Subramaniyan Shanmuganathan, S.R. Nagasupreeta, Manchikanti Sri Sai Charan, Ganesh Sheshagiri, Vivek Gairola, Suresh H. Basagoudanavar , B.P. Sreenivasa , Veerakyathappa Bhanuprakash
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引用次数: 0
摘要
在口蹄疫(FMD)流行的国家,定期接种疫苗是控制疾病发病率的关键手段。对接种疫苗的人群进行积极的血清监测对于评估疫苗接种的效果至关重要。病毒中和试验(VNT)和酶联免疫吸附试验(ELISA)常用于抗体检测。液相阻断酶联免疫吸附试验(LPBE)或固相竞争酶联免疫吸附试验(SPCE)是首选的检测方法,因为它们不需要处理口蹄疫病毒,可常规用于血清监测或疫苗效力测试;但是,LPBE 的假阳性率很高。在此,我们报告了一种基于单克隆抗体(mAb)的 SPCE,作为抗体滴定的潜在替代检测方法。从 12 种针对 FMDV 血清型 A 的 mAb 中,我们选择了两种 mAb 用于 SPCE 的开发。结果表明,基于 mAb 2C4G11、mAb 6E8D11 和 pAb 的 SPCE 对 453 份血清的相对灵敏度分别为 86.1%、86.1% 和 80.3%,特异性分别为 99.6%、99.1% 和 99.1%。上述检测方法的相关性、可重复性和一致程度都很高,这表明 mAb 可用于大规模监测研究和常规疫苗效力检测。
Monoclonal antibody based solid phase competition ELISA to detect FMDV serotype A specific antibodies
In Foot-and-mouth disease (FMD) enzootic countries, periodic vaccination is the key tool in controlling the disease incidence. Active seromonitoring of the vaccinated population is critical to assess the impact of vaccination. Virus neutralization test (VNT) and enzyme-linked immunosorbent assays (ELISA) are commonly used for antibody detection. Assays like liquid phase blocking ELISA (LPBE) or solid phase competition ELISA (SPCE) are preferred as they do not require handling of live FMDV and are routinely used for seromonitoring or for vaccine potency testing; however, false positives are high in LPBE. Here we report, a monoclonal antibody (mAb) based SPCE as a potential alternate assay for antibody titration. From a panel of 12 mAbs against FMDV serotype A, two mAbs were chosen for the development of SPCE. Based on a set of 453 sera, it was demonstrated that mAb 2C4G11, mAb 6E8D11and polyclonal antibody (pAb) based SPCE had a relative sensitivity of 86.1, 86.1 and 80.3 %; and specificity of 99.6, 99.1 and 99.1 %, respectively. The correlation, repeatability, and level of agreement of the assays were high demonstrating the potential use of mAb in large scale surveillance studies and regular vaccine potency testing.
期刊介绍:
The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery.
The methods may include, but not limited to, the study of:
Viral components and morphology-
Virus isolation, propagation and development of viral vectors-
Viral pathogenesis, oncogenesis, vaccines and antivirals-
Virus replication, host-pathogen interactions and responses-
Virus transmission, prevention, control and treatment-
Viral metagenomics and virome-
Virus ecology, adaption and evolution-
Applied virology such as nanotechnology-
Viral diagnosis with novelty and comprehensive evaluation.
We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.