Multicolor Immunodiagnostic for Semiquantitative Visual Detection Syndrome Coronavirus 2 Specific Antibody： A Prospect Strategy for On-site COVID-19 Therapeutic Process Monitoring

doi:10.7503/cjcu20210363

Abstract

Abstract:

Rapid detection of body fluid severe acute respiratory syndrome coronavirus 2（SARS-CoV-2） antibody is an effective strategy for infection therapeutic effect of coronavirus disease（COVID-19）. Most detection methods require relatively large equipment， which limited their on-site application. Lateral flow immunoassay（LFIA） can be used to qualitative antibody detection based on the aggregation of gold nanoparticles （Au NPs）， which exhibits just one-color change and cannot realize rapid quantitative detection without the help of additional equipment. In this study， a high-resolution multicolor colorimetric strategy was developed and applied to assessing antibody concentration at a glance based on etching of gold nanorods（Au NRs）. Firstly， SARS-CoV-2 recombinant antigen was immobilized on the surface of the 96-wells. Then， horseradish peroxidase（HRP）-labeled second antibody combined with antibody to form an antigen-antibody-secondary antibody complex on the well surface， which has direct relationship with antibody concentration in the sample and can be used to oxidize 3，3′，5，5′-tetramethylbenzidine（TMB） to form TMB²⁺ at the presence of HRP. The generation of TMB²⁺ efficiently etch Au NRs to produce multicolor solution. The etching result in vivid color changes in the system has a relationship with the amount of SARS-CoV-2 IgM antibody. Under the optimal conditions， the proposed strategy exhibited a linear response in the 5.00―200 IU concentration range， and a detection limit of 1.29 IU for SARS-CoV-2 IgM antibody， with high sensitivity and specificity. This assay is prospective for the on-site semi-quantitative visual detection of SARS-CoV-2 IgM antibody concentration in the COVID-19 therapeutic process.

Key words: Severe acute respiratory syndrome coronavirus 2（SARS-CoV-2）, Antibody, Multicolor colorime-tric strategy, Therapeutic process monitoring

CLC Number:

O657

TrendMD:

CHEN Zhonghui, LI Jinqiu, LIN Wei, YU Liumin, TU Haijian, CHEN Yu, CAI Zongwei, LIN Zhenyu. Multicolor Immunodiagnostic for Semiquantitative Visual Detection Syndrome Coronavirus 2 Specific Antibody： A Prospect Strategy for On-site COVID-19 Therapeutic Process Monitoring[J]. Chem. J. Chinese Universities, 2021, 42(11): 3509.

Figures/Tables 6

Fig.1 UV?Vis spectra(A―D) and corresponding TEM images(A′―D′) of Au NRs with addition of different concentrations of SARS?CoV?2 IgM antibodyIgM antibody concentration/IU: (A, A′) 0; (B, B′) 10.0; (C, C′) 60.0; (D, D′) 100. Insets in (A)―(D) show the relative solution color change.

Fig.2 Effects of CTAB concentration(A), HCl concentration(B), the HRP?Ab capture reaction time(C) and TMB liquid substrate system reaction time(D) on the LSPR peak blue shift of Au NRs

Fig.3 Plasmonic method color change with SARS?CoV?2 antibody concentration increased(A), UV?Vis spectra change of Au NRs in the oxidation etching process with different SARS?CoV?2 antibody concentrations presented(B) and the linear relationship between Δλ and SARS?CoV?2 antibody concentration(C)The SARS?CoV?2 concentrations of a―h(A) and a―h(B)/IU: 0, 1.00, 5.00, 10.0, 30.0, 60.0, 100, 200.

Fig.4 Selectivity of the developed multicolor assay detection of SARS?CoV?2 IgM antibodya, a′. Target; b, b′. blank; c, c′. H5N1Ag; d, d′. H7N9Ag; e, e′. HBsAb; f, f′. HCVAb. Error bars represent the standard error derived from three repeated measurement. The inset images exhibit the color of the corresponding solutions.

Table 1 Detection of SARS-CoV-2 IgM antibody between healthy volunteers and COVID-19 patients by the proposed method

Sample source	Detected concentration/（IU， mean±SD）	RT?PCR result
Healthy volunteer 1	0.53±0.16	Negative
Healthy volunteer 2	0.71±0.08	Negative
Healthy volunteer 3	0.28±0.12	Negative
COVID?19 patient 1	17.3±2.8	Positive
COVID?19 patient 2	6.30±1.41	Positive
COVID?19 patient 3	26.8±3.0	Positive

Table 2 Detection of a typical COVID-19 patient SARS-CoV-2 IgM antibody concentration change in therapeutic and convalescent processes by the proposed method

Days after symptom onset	Detected concentration/（IU， Mean±SD）	LFIA result
5	2.95±0.81	+/-
8	7.43±1.39	++
12	27.3±2.4	+++
20	44.43±3.6	+++
30	22.3±2.2	+++
45	14.7±1.1	++
60	3.87±0.55	+

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