Abstract
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.
Highlights
Between 2007 and 2010, a severe febrile illness was associated with gastrointestinal symptoms, thrombocytopenia, leukocytopenia, and high mortality in the eastern and central regions of China [1,2].This disease is called severe fever with thrombocytopenia syndrome (SFTS) and was caused by the newly discovered bunyavirus (SFTSV) [3]
The results described in these prior studies all prove that AuNPs have a large capacity for carrying proteins—this is a great advantage in the field of improving detection sensitivity
In the indirect enzyme-linked immunosorbent assay (ELISA) test, the antibody in the sample was sandwiched between the antigen coated on the 96-well plate and the enzyme-labeled secondary antibody
Summary
Between 2007 and 2010, a severe febrile illness was associated with gastrointestinal symptoms, thrombocytopenia, leukocytopenia, and high mortality in the eastern and central regions of China [1,2]. There is an urgent need to develop ultrasensitive detection methods for the different types of biomarkers In this context, nanotechnology offers many ways to improve detection sensitivity. AuNPs are distinguished from other nanoparticles and quantum dots containing harmful heavy metal ions because of their simpler synthesis process and effective surface modification [22] Their high surface area can carry many biomolecules (such as antibodies, enzymes, or DNA) to produce significant signal enhancement [23,24]. Jia [26] and Wu [27] developed a dual-modified gold nanoprobes for enhanced immunoassay using the same experimental principle In their experiments, they used AuNPs as a bridge between the detection antibody and HRP. The enzyme freely moved in solution, which allowing the substrate to be close to the enzyme without encountering steric hindrance from the AuNPs
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