气候变化领域集成服务门户(CCPortal): Direct single-molecule imaging for diagnostic and blood screening assays

CCPortal

DOI	10.1073/pnas.2025033118
	Direct single-molecule imaging for diagnostic and blood screening assays
	Ruan Q.; Macdonald P.J.; Swift K.M.; Tetin S.Y.
发表日期	2021
ISSN	00278424
卷号	118 期号:14
英文摘要	Every year, over 100 million units of donated blood undergo mandatory screening for HIV, hepatitis B, hepatitis C, and syphilis worldwide. Often, donated blood is also screened for human T cell leukemia-lymphoma virus, Chagas, dengue, Babesia, cytomegalovirus, malaria, and other infections. Several billion diagnostic tests are performed annually around the world to measure more than 400 biomarkers for cardiac, cancer, infectious, and other diseases. Considering such volumes, every improvement in assay performance and/or throughput has a major impact. Here, we show that medically relevant assay sensitivities and specificities can be fundamentally improved by direct single-molecule imaging using regular epifluorescence microscopes. In current microparticle-based assays, an ensemble of bound signal-generating molecules is measured as a whole. By contrast, we acquire intensity profiles to identify and then count individual fluorescent complexes bound to targets on antibody-coated microparticles. This increases the signal-to-noise ratio and provides better discrimination over nonspecific effects. It brings the detection sensitivity down to the attomolar (10-18 M) for model assay systems and to the low femtomolar (10-16 M) for measuring analyte in human plasma. Transitioning from counting single-molecule peaks to averaging pixel intensities at higher analyte concentrations enables a continuous linear response from 10-18 to 10-5 M. Additionally, our assays are insensitive to microparticle number and volume variations during the binding reaction, eliminating the main source of uncertainties in standard assays. Altogether, these features allow for increased assay sensitivity, wide linear detection ranges, shorter incubation times, simpler assay protocols, and minimal reagent consumption. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Diagnostics; Fluorescence; Immunoassays; Microparticles; Single-molecule imaging
语种	英语
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/179978
作者单位	Applied Research and Technology, Abbott Diagnostics Division, Abbott Laboratories, Abbott Park, IL 60064, United States
推荐引用方式 GB/T 7714	Ruan Q.,Macdonald P.J.,Swift K.M.,et al. Direct single-molecule imaging for diagnostic and blood screening assays[J],2021,118(14).
APA	Ruan Q.,Macdonald P.J.,Swift K.M.,&Tetin S.Y..(2021).Direct single-molecule imaging for diagnostic and blood screening assays.Proceedings of the National Academy of Sciences of the United States of America,118(14).
MLA	Ruan Q.,et al."Direct single-molecule imaging for diagnostic and blood screening assays".Proceedings of the National Academy of Sciences of the United States of America 118.14(2021).