Highly Sensitive Fluorescence-Linked Immunosorbent Assay for the Determination of Human IgG in Serum Using Quantum Dot Nanobeads and Magnetic Fe(3)O(4) Nanospheres #MMPMID32954173
Guo J; Wang Y; Niu S; Li H; Tian Y; Yu S; Yu F; Wu Y; Liu LE
ACS Omega 2020[Sep]; 5 (36): 23229-23236 PMID32954173show ga
The aim of this study is to establish a new method with high sensitivity, accuracy, and stability for the determination of human IgG and then expand it to analyze severe acute respiratory syndrome corona virus 2 (SARS-CoV-2)-specific IgM and IgG, which is of great significance for the screening and diagnosis of COVID-19. In this study, the magnetic Fe(3)O(4) nanospheres coupled with mouse antihuman IgG (Ab1(IgG)) were used as an immune capture probe (Fe(3)O(4)@Ab1(IgG)) to capture and separate the target, and rabbit antihuman IgG (Ab2(IgG)) coupled with highly luminescent quantum dot nanobeads (QBs) as a fluorescence detection probe (QBs@Ab2(IgG)) was used to realize high sensitivity detection. After the formation of a sandwich immunocomplex, the fluorescence intensity of the precipitate after magnetic separation was measured at the excitation wavelength of 370 nm. Under optimal conditions, a wide linear range varying from 0.005 to 40 ng.mL(-1) was obtained for the detection of human IgG with a lower limit of detection at 4 pg.mL(-1) (S/N = 3). The recoveries of intra- and interassays were 90.0-101.9 and 96.0-106.6%, respectively, and the relative standard deviations were 6.3-10.2 and 2.6-10.5%, respectively. Furthermore, the proposed method was successfully demonstrated to detect human IgG in serum samples, and the detection results were not statistically different (P > 0.05) from commercial enzyme-linked immunosorbent assay kits. This method is sensitive, fast, and accurate, which could be expanded to detect the specific IgM and IgG antibodies against SARS-CoV-2.
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ACS+Omega 2020 ; 5 (36): 23229-23236
