CN-115353557-B - MOFs nano-enzyme and application thereof in detection of novel coronaviruses

Abstract

A MOFs nano-enzyme and application thereof in detection of new coronaviruses relate to the field of biological detection methods, and the MOFs nano-enzyme has a peroxidase-like effect and can catalyze a color developing agent to change color, after a specific protein CD147 is modified on the surface of the nano-enzyme, the specific combination of the CD147 and an S protein over-expressed on the surface of the new coronaviruses is utilized to efficiently combine the new coronaviruses on the surface of the nano-enzyme so as to inhibit the catalytic color development of the nano-enzyme, and based on the principle, the quantitative and qualitative detection of the new coronaviruses by a colorimetric method can be realized.

Inventors

• MENG LINGJIE
• ZHAO XIAOPING
• YANG ZHIWEI

Assignees

• 西安交通大学

Dates

Publication Date

20260508

Application Date

20220822

Claims (2)

1. 1. The application of MOFs nano-enzyme in detection of novel coronaviruses for non-disease diagnosis is characterized in that the specific binding capacity of MOFs enzyme-like characteristics and surface modification CD147 proteins to the novel coronaviruses can catalyze the molecular chromogenic capacity of chromogenic molecules TMB, ABTS, OPD and PPD, and the colorimetric quantitative and qualitative detection of SARS-CoV-2 is realized; The MOFs nano enzyme is a MOFs material of Fe and Cu, a microwave-assisted solvothermal method is adopted, terephthalic acid is used as a ligand, metal ions are chelated with the terephthalic acid to form a metal MOFs material, carboxyl groups are arranged on the surface of the metal MOFs material, and then the metal MOFs material is condensed with CD147 molecules with amino groups to fix the CD147 molecules on the surface of the MOFs, wherein the proportion of the ligand to the metal ions is (1-10), the proportion of the Fe and the Cu ions is (1-50), the reaction temperature is 120-160 ℃, and the reaction time is 2-12h; The MOFs nano enzyme has a peroxidase-like characteristic, can catalyze hydrogen peroxide to generate hydroxyl free radicals, oxidize chromogenic molecules TMB, ABTS, OPD and PPD, and realize detection of the novel coronavirus by a colorimetric method; TMB is 3,3', 5' -tetramethyl lbenzidine; ABT is 2,2' -azinobis- (3-ethyllbenzthiazoline-6-sulfate); OPD is o-PHENYLENEDIAMINE; PPD is p-PHENYLENEDIAMINE; The MOFs nano-enzyme is modified with a specific protein CD147 on the surface, and the specific combination of the CD147 and the S protein over-expressed on the surface of the novel coronavirus SARS-CoV-2 is utilized to combine the novel coronavirus on the surface of the nano-enzyme so as to inhibit the catalytic color development of the nano-enzyme, thereby realizing the quantitative qualitative analysis of the novel coronavirus SARS-CoV-2; after modifying CD147 by MOFs nano-enzyme, the catalytic activity of MOFs is reduced, the capacities of oxidizing TMB, ABTS, OPD and changing color of PPD molecules are reduced, and the modified CD147 is formed by condensation connection of carboxyl on the surface of MOFs nano-enzyme and amino of CD 147.
2. 2. The use of MOFs nanoenzyme according to claim 1, wherein the CD147 specifically binds to SARS-CoV-2, the catalytic activity of MOFs nanoenzyme is reduced, and the ability to oxidize TMB, ABTS, OPD and PPD molecules to change color is further weakened.

Description

MOFs nano-enzyme and application thereof in detection of novel coronaviruses Technical Field The invention belongs to the technical field of biological medicine and clinical diagnosis, in particular relates to MOFs nano-enzyme and application thereof in new coronavirus detection, and specifically relates to a method for realizing visual, rapid and high-sensitivity detection of SARS-CoV-2 by using the MOFs nano-enzyme. Background The novel coronavirus 2019-nCoV/SARS-CoV-2 is in the world-wide momentum, but the human beings lack effective antiviral vaccines and specific drugs, and the accurate detection and diagnosis of the infected person or carrier is of great importance to epidemic control. Currently, COVID-19 diagnostic strategies are based mainly on three methods, nucleic acid detection, antibody detection and antigen detection. Nucleic acid detection is a "gold standard" for clinical diagnosis of new coronaries, and low amounts of virus are detected based on nucleic acid amplification. Reverse transcription polymerase chain reaction (RT-PCR) in combination with an automated sample preparation system allows for the bulk detection of SARS-CoV-2 nucleic acid present in the nasopharynx or saliva. However, the detection result of nucleic acid is affected by the factors such as the course of the patient, sample collection, quality of detection reagent and the like, and the positive detection rate is only 30-50%. Therefore, improvement of detection sensitivity and specificity is still an urgent need to be solved for improvement of detection sensitivity and efficiency. In addition, expensive and inflexible detection equipment also limits its use in large scale multi-sample applications. Antibody detection, which is primarily the detection of novel coronavirus specific IgM and IgG antibodies in serum, is a complementary and rapid method of viral detection, but is generally applicable to those who are only diagnosed for at least 10-14 days after infection, which may lead to false negative results. In order to cope with these problems, in recent years, various detection methods based on specific viral proteins (including antigens) have been proposed for identifying the presence of pathogens due to their high sensitivity, high selectivity and rapid measurement, and the probability of cross-reactivity can be reduced, effectively improving the specificity thereof. Among these methods, colorimetric methods are attractive virus detection methods because of their simplicity, economy, and particular ease of visual readout, and such assays have great potential to be performed at remote sites and can be used without the need for complex equipment. Enzyme catalysis is one of the most commonly used colorimetry methods for catalyzing the color change of chromogenic molecules, however, the enzyme is easy to deactivate under severe conditions, and the application of the enzyme is limited. In recent years, metal-Organic Framework Structures (MOFs) have been widely used in the construction of colorimetric biosensors because they have the same function as peroxidases and can decompose H 2O2 to produce OH and develop color with an oxidation developer. Disclosure of Invention In order to overcome the defects of the existing novel coronal detection method, the invention aims to provide MOFs nano-enzyme and application thereof in novel coronavirus detection, and is a novel method for simply, rapidly, highly sensitively and label-free visual detection of SARS-CoV-2 based on MOFs nano-enzyme. In order to achieve the above object, the present invention adopts the following technical scheme: a MOFs nano-enzyme is prepared from terephthalic acid as ligand, and metal ions through chelating to form metallic MOFs material, which has carboxyl on its surface, and the condensation of CD147 molecule with amino group to make CD147 molecule fixed to MOFs surface, and one or more of Fe, cu, ni and Pt. Based on the application of MOFs nano-enzyme in detection of new coronaviruses, the specific binding capacity of MOFs enzyme-like characteristics and surface modification CD147 proteins to the new coronaviruses can catalyze the molecular chromogenic capacity of chromogenic molecules TMB, ABTS, OPD and PPD, and the colorimetric quantitative and qualitative detection of SARS-CoV-2 can be realized. The MOFs nano-enzyme has peroxidase-like characteristics, can catalyze hydrogen peroxide to generate hydroxyl free radicals, oxidize chromogenic molecules TMB, ABTS, OPD and PPD, and realize detection of the novel coronavirus by a colorimetric method. TMB is 3,3', 5' -tetramethyl lbenzidine, ABT is 2,2' -azinobis- (3-ethyllbenzylthiazoline-6-sulfophone), OPD is o-PHENYLENEDIAMINE, PPD is p-PHENYLENEDIAMINE. The MOFs nano-enzyme is modified with a specific protein CD147 on the surface, and the specific combination of the CD147 and the S protein over-expressed on the surface of the novel coronavirus SARS-CoV-2 is utilized to combine the novel coronavirus on the s