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Enzyme-Antibody-Modified Gold Nanoparticle Probes for the Ultrasensitive Detection of Nucleocapsid Protein in SFTSV

机译:酶 - 抗体改性的金纳米颗粒探头,用于SFTSV中核衣壳蛋白的超细囊炎

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摘要

As humans and climate change continue to alter the landscape, novel disease risk scenarios have emerged. Sever fever with thrombocytopenia syndrome (SFTS), an emerging tick-borne infectious disease first discovered in rural areas of central China in 2009, is caused by a novel bunyavirus (SFTSV). The potential for SFTS to spread to other countries in combination with its high fatality rate, possible human-to-human transmission, and extensive prevalence among residents and domesticated animals in endemic regions make the disease a severe threat to public health. Because of the lack of preventive vaccines or useful antiviral drugs, diagnosis of SFTS is the key to prevention and control of the SFTSV infection. The development of serological detection methods will greatly improve our understanding of SFTSV ecology and host tropism. We describe a highly sensitive protein detection method based on gold nanoparticles (AuNPs) and enzyme-linked immunosorbent assay (ELISA)—AuNP-based ELISA. The optical sensitivity enhancement of this method is due to the high loading efficiency of AuNPs to McAb. This enhances the concentration of the HRP enzyme in each immune sandwich structure. The detection limit of this method to the nucleocapsid protein (NP) of SFTSV was 0.9 pg mL−1 with good specificity and reproducibility. The sensitivity of AuNP-based ELISA was higher than that of traditional ELISA and was comparable to real-time quantitative polymerase chain reaction (qRT-PCR). The probes are stable for 120 days at 4 °C. This can be applied to diagnosis and hopefully can be developed into a commercial ELISA kit. The ultrasensitive detection of SFTSV will increase our understanding of the distribution and spread of SFTSV, thus helping to monitor the changes in tick-borne pathogen SFTSV risk in the environment.
机译:随着人类和气候变化继续改变景观,出现了新的疾病风险情景。 2009年中国中美洲农村首次发现的新出现的蜱型传染病综合征(SFT)的切除发烧是由新的Bunyavirus(SFTSV)引起的。 SFT的潜力与其具有高死亡率,可能的人对人类传播的潜力,以及人类对人类的人类传播,流行区域中的居民和驯养动物的广泛普遍使疾病对公共卫生的严重威胁。由于缺乏预防疫苗或有用的抗病毒药物,SFT的诊断是预防和控制SFTSV感染的关键。血清学检测方法的发展将大大提高我们对SFTSV生态和主持人的理解。我们描述了一种基于金纳米颗粒(AUNP)和酶联免疫吸附测定(ELISA)-AUNP的ELISA的高灵敏的蛋白质检测方法。该方法的光学灵敏度增强是由于AUNPS到MCAB的高负载效率。这增强了每个免疫夹层结构中HRP酶的浓度。该方法对SFTSV核衣壳蛋白(NP)的检测限为0.9pg ml-1,具有良好的特异性和再现性。基于AUNP的ELISA的敏感性高于传统ELISA的敏感性,与实时定量聚合酶链反应(QRT-PCR)相当。探针在4°C下稳定为120天。这可以应用于诊断,并希望可以开发成商业ELISA套件。 SFTSV的超敏检测将增加我们对SFTSV的分布和传播的理解,从而有助于监测环境中蜱传播病原体SFTSV风险的变化。

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