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WO2021184391A1 - Procédé de criblage précoce du nouveau coronavirus - Google Patents

Procédé de criblage précoce du nouveau coronavirus Download PDF

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Publication number
WO2021184391A1
WO2021184391A1 PCT/CN2020/080524 CN2020080524W WO2021184391A1 WO 2021184391 A1 WO2021184391 A1 WO 2021184391A1 CN 2020080524 W CN2020080524 W CN 2020080524W WO 2021184391 A1 WO2021184391 A1 WO 2021184391A1
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cov
sars
protein
kit
iga
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Chinese (zh)
Inventor
金腾川
姜德华
马欢
常志远
曾威红
易如婷
黄文喜
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Guangzhou Kangrun Biotechnology Co Ltd
Bioisland Laboratory
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Guangzhou Kangrun Biotechnology Co Ltd
Bioisland Laboratory
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/62—DNA sequences coding for fusion proteins
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • C12N7/04—Inactivation or attenuation; Producing viral sub-units
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A61K39/215—Coronaviridae, e.g. avian infectious bronchitis virus

Definitions

  • the present invention relates to the technical field of immunoassays, in particular to an earlier screening method, reagents and kits for SARS-CoV-2 IgA antibodies of novel coronavirus infection pneumonia (new coronary pneumonia).
  • coronavirus pneumonia Since December 2019, the new type of coronavirus pneumonia (new crown pneumonia) has been raging. The cumulative number of people infected in China has exceeded 80,000, and the mortality rate has reached 1.4-3.4%.
  • New coronary pneumonia usually develops 3-7 days after being infected with SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2). The symptoms are mainly fever, dry cough, fatigue, and can also be accompanied by nasal congestion, runny nose, sore throat, myalgia and diarrhea, etc. . Mild patients have no symptoms of pneumonia. Severe patients often develop dyspnea and/or hypoxemia one week after the onset of onset.
  • the virus nucleic acid detection reagent was first used for the diagnosis of new coronary pneumonia. Because of its low detection rate, many companies have begun to develop rapid detection kits for SARS-CoV-2IgG and IgM, including colloidal gold method and chemiluminescence immunoassay. The seventh edition of the new coronary pneumonia diagnosis and treatment plan incorporates IgM and IgG serological testing as laboratory diagnostic methods. Currently, the registered products include 11 nucleic acid detection products and 6 antibody detection products.
  • the latter include: colloidal gold method new coronavirus (2019-nCoV) antibody detection kit, colloidal gold method new coronavirus (2019-nCoV) IgM Antibody detection kit, magnetic particle chemiluminescence method new coronavirus (2019-nCoV) IgM antibody detection kit, magnetic particle chemiluminescence method new coronavirus (2019-nCoV) IgG antibody detection kit, new type of scientific luminescent particle immunoassay method Coronavirus (2019-nCoV) antibody detection kit.
  • SARS-CoV-2 infects the lower respiratory tract, the patient produces less sputum, and there is a high probability that the sample collection will fail with a throat swab.
  • the detection rate of nucleic acid detection reagents for new coronary pneumonia is low, and the sensitivity is only 30-50%, which cannot meet the needs of screening all patients.
  • IgM and IgG antibodies Measuring the total antibody improves the sensitivity but cannot effectively distinguish which stage of the patient is in the course of the disease and whether it is still infectious.
  • the colloidal gold method is simple and quick to detect, but the accuracy is not enough.
  • the sensitivity and specificity of chemiluminescence detection of IgM and IgG antibodies are high, but the window period of the two is long. IgM needs to be detected 3-5 days after the onset of onset, and the detection value of IgG needs to increase by more than 4 times during the recovery period to be diagnosed.
  • the purpose of the present invention is to avoid the shortcomings of the prior art and provide a kit and reagents for screening new coronavirus infections that can detect SARS-CoV-2 IgA antibodies earlier.
  • kits for detecting SARS-CoV-2 IgA antibody for screening new coronavirus infection Provide a kit for detecting SARS-CoV-2 IgA antibody for screening new coronavirus infection.
  • the above-mentioned kit for screening for novel coronavirus infection is used for early screening of SARS-CoV-2 IgA antibodies.
  • the above kit for screening for novel coronavirus infection is used as the SARS-CoV-2 magnetic bead coating of R1: a magnetic bead coating containing SARS-CoV-2 recombinant antigen-coated trimethylol Aminomethane buffer, pH value is 7.1-7.4;
  • R2 anti-human IgA antibody acridinium ester label 2-(N-morpholine) ethanesulfonic acid buffer containing acridinium ester-labeled mouse anti-human IgA monoclonal antibody.
  • the SARS-CoV-2 recombinant antigen is a spike protein expressed in a suspension cultured human embryonic kidney cell line HEK293F, referred to as S protein (Spike protein, S protein). ), the amino acid sequence at positions Gln321-Ser591 of the RBD domain was truncated for protein expression;
  • the protein expression process is: the SARS-CoV-2 S protein RBD gene is connected to human IgG1 Fc through a TEV restriction site sequence at the C-terminus, and then it is cloned into the mammalian expression vector pTT5 and Kozak is added to the translation initiation region The sequence was then transfected into suspension culture cells HEK293F, using interferon ⁇ -1 as a signal peptide to make HEK293F cells secrete RBD-TEV-Fc fusion protein, and then using Protein A column and nickel column for two purifications; the first purification To use Protein A column to obtain RBD-TEV-Fc fusion protein from the culture supernatant; the second purification is to use TEV enzyme with 6His tag to digest the fusion protein, and then pass the digested products through Protein A column in turn And nickel column to remove Fc, TEV enzymes, and the impurity protein bound to the Protein A column during the first purification, and finally obtain pure RBD protein in the flow-
  • the SARS-CoV-2 recombinant antigen is a nucleocapsid protein expressed in bacteria in suspension culture, referred to as N protein, and N protein is a cut of Met1-Ala419 The amino acid sequence at position is expressed;
  • the protein expression process is: connect the SARS-CoV-2 N protein gene to the 6His tag at the N end and clone it into the prokaryotic expression vector pET28a, then transform it into E. coli cells BL21 for N protein expression, and then take the broken bacteria and lyse the bacteria. Add 0.5-2.0M sodium chloride to the solution to reduce non-specific binding of protein and nucleic acid, and then pass the lysate through a nickel column.
  • the tris buffer contains 0.05-0.2% Tween 20 (v/v), 2-8% bovine serum albumin ( w/v), 0.25-1% Proclin300(v/v);
  • 2-(N-morpholine) ethanesulfonic acid buffer contains 0.05-0.2% Tween 20 (v/v), 2-8% bovine serum albumin (w/v), 0.25-1% Proclin 300 (v/v) ).
  • the above kit for screening for novel coronavirus infection also contains:
  • reaction buffer of R3 phosphate buffer containing goat anti-human IgG antibody, containing 0.05-0.2% Tween 20 (v/v), 2-8% bovine serum albumin (w/v), 0.25- 1% Proclin300(v/v);
  • SARS-CoV-2 IgA negative control SARS-CoV-2 IgA negative serum
  • SARS-CoV-2 IgA positive control human-derived purified SARS-CoV-2 IgM antibody
  • Negative and positive control information card contains the luminescence value of the negative and positive control, which is used for correction during screening.
  • the above-mentioned kit for screening for novel coronavirus infection contains:
  • Test card It is composed of SARS-CoV-2 IgA test strip and plastic box; the test strip contains nitrocellulose membrane, sample pad, bonding pad, absorbent paper, and PVC board support;
  • the nitrocellulose membrane is coated with anti-human-IgA antibody and anti-mouse IgG polyclonal antibody, and the binding pad contains colloidal gold-labeled SARS-CoV-2 antigen.
  • the above-mentioned kit for screening for novel coronavirus infection also contains Tris hydrochloric acid buffer as a sample diluent, the pH of the Tris hydrochloric acid buffer is 7.1-7.4, and it contains 0.5-2% bovine serum white Protein (w/v), 0.05-0.2% Tween 20 (v/v), 0.25-1% Proclin 300 (v/v).
  • the present invention also provides a use of a reagent for detecting SARS-CoV-2 IgA antibody in preparing the above reagent or kit for screening novel coronavirus infection.
  • the kit used for screening the novel coronavirus infection and its detection reagent of the present invention can detect the novel coronavirus.
  • Figure 1 is a schematic diagram of the results of serum SARS-CoV-2 IgA antibody levels in patients with new coronary pneumonia and a control group in an experimental example of the present invention.
  • Fig. 2 is a graph showing the change trend of SARS-CoV-2 IgA and IgM antibodies with the course of the disease in an experimental example of the present invention.
  • a kit for the detection of SARS-CoV-2 IgA antibodies for screening new coronavirus infections for screening new coronavirus infections. Especially for early screening of SARS-CoV-2 IgA antibodies.
  • This kit for screening for novel coronavirus infection includes:
  • SARS-CoV-2 magnetic bead coating (R1): Tris buffer containing magnetic bead coating of SARS-CoV-2 recombinant antigen, pH 7.1-7.4, containing 0.05 -0.2% Tween 20 (v/v), 2-8% bovine serum albumin (w/v), 0.25-1% Proclin 300 (v/v); magnetic beads are used as the reaction solid phase carrier, and the antigen is used for binding The target antibody in the test substance.
  • Anti-human IgA antibody acridinium ester marker (R2): 2-(N-morpholine) ethanesulfonic acid buffer of acridinium ester-labeled mouse anti-human IgA monoclonal antibody, containing 0.05-0.2% Tween 20 (v/v), 2-8% bovine serum albumin (w/v), 0.25-1% Proclin300 (v/v). Combining with the antigen-antibody complex, under alkaline conditions, the acridinium ester can emit light after being excited.
  • Reaction buffer (R3) phosphate buffer containing goat anti-human IgG antibody, containing 0.05-0.2% Tween 20 (v/v), 2-8% bovine serum albumin (w/v), 0.25-1% Proclin300 (v/v). Used to adsorb IgG in the sample to be tested to prevent interference.
  • SARS-CoV-2 IgA negative control SARS-CoV-2 IgA negative serum.
  • SARS-CoV-2 IgA positive control It is a human-derived purified SARS-CoV-2 IgM antibody.
  • Negative and positive control information card contains the luminescence value of the negative and positive control, which is used for correction during screening.
  • the experiment requires the substrate solution (aqueous solution containing NaOH) and the cleaning solution (phosphate buffer containing surfactant) as general reagents, which are not provided in this kit.
  • SARS-CoV-2 IgA turns positive earlier than IgM antibodies, 1-4 days earlier on average. Therefore, the clinical detection of SARS-CoV-2 IgA antibodies can be carried out earlier, and some patients with new coronary pneumonia can be screened and dealt with early.
  • SARS-CoV-2 recombinant antigen is in Spike protein (S protein) expressed in suspension cultured human embryonic kidney cell line HEK293F, referred to as S protein, was extracted from the Gln321-Ser591 amino acid sequence of the RBD domain for protein expression.
  • S protein Spike protein
  • the protein expression process is: the SARS-CoV-2 S protein RBD gene is connected to human IgG1 Fc through a TEV restriction site sequence at the C-terminus, and then it is cloned into the mammalian expression vector pTT5 and Kozak is added to the translation initiation region The sequence was then transfected into suspension culture cells HEK293F, using interferon ⁇ -1 as a signal peptide to make HEK293F cells secrete RBD-TEV-Fc fusion protein, and then using Protein A column and nickel column for two purifications; the first purification To obtain the RBD-TEV-Fc fusion protein from the culture supernatant using the Protein A column; the second purification is to digest the fusion protein with the TEV enzyme with 6His tag, and then flow the digested products through the Protein A column in turn And nickel column to remove Fc, TEV enzymes, and the impurity protein bound to the Protein A column during the first purification, and finally obtain pure RBD protein in the
  • the SARS-COV-2 spike RBD protein expressed by this method is intercepted from the Gln321-Ser591 amino acid sequence of the SARS-COV-2 spike protein.
  • the coding gene sequence is shown in the following sequence 1, and its amino acid sequence is shown in the following sequence 2.
  • Sequence 1 The gene sequence encoding SARS-COV-2 spike RBD in this patent
  • the SARS-COV-2 spike RBD protein of this method is secreted and expressed, and the signal peptide used is the IFNA1 (Interferon ⁇ -1) signal peptide, and the Kozak sequence is added to the translation initiation region.
  • the C-terminus of the SARS-COV-2 spike RBD protein of this patent is connected to the human IgG1 Fc sequence through a TEV restriction site sequence.
  • the expressed fusion protein is named SARS-COV-2-RBD-TEV-Fc, and its gene sequence (including The signal peptide) is shown in the following sequence 3, and the amino acid sequence is shown in the following sequence 4.
  • Sequence 3 The gene sequence of the fusion protein SARS-COV-2-RBD-TEV-Fc
  • This patent obtains high-purity SARS-COV-2 spike RBD protein through two purifications.
  • the protein A column was used to obtain the SARS-COV-2-RBD-TEV-Fc fusion protein from the culture supernatant.
  • the fusion protein was digested with TEV enzyme with 6His tag, and the digested products were passed through the protein A column and the nickel column in turn. At this time, the Fc, TEV enzyme, and the protein bound to the Protein A column during the first purification were all removed. After removal, the purified SARS-COV-2 spike RBD protein is finally obtained in the flow through.
  • the SARS-CoV-2 recombinant antigen is a nucleocapsid protein expressed in bacteria in suspension culture, referred to as the N protein, and the N protein is expressed by truncating the amino acid sequence at position Met1-Ala 419.
  • the protein expression process is: connect the SARS-CoV-2 N protein gene to the 6His tag at the N end and clone it into the prokaryotic expression vector pET28a, then transform it into E. coli cells BL21 for N protein expression, and then take the broken bacteria and lyse the bacteria.
  • the SARS-COV-2 N protein expressed in this patent is truncated from the SARS-COV-2 N protein Met1-Ala 419 amino acid sequence.
  • the coding gene sequence is shown in the following sequence 5, and its amino acid sequence is shown in the following sequence 6.
  • the SARS-COV-2 N protein of this patent is soluble expression in bacteria, with 6His tag connected to its N end, and a nickel column is used to purify the soluble SARS-COV-2 N from the supernatant of the bacterial fragments. At this time, it is purified. A large amount of nucleic acid is bound to the N protein. In order to remove the nucleic acid, ammonium sulfate is added and stirred at room temperature to expose the hydrophobic core of the protein, and then a hydrophobic column is used to further purify the protein. At this time, the nucleic acid is removed. In order to further remove polymers and other impurity proteins, the superdex200 molecular sieve was used to further purify the protein, and finally a high-purity SARS-COV-2 N protein with uniform conformation was obtained.
  • the kit of the present invention can detect the SARS-CoV-2 IgA antibody through the clinic earlier, and screen and deal with some patients with new coronary pneumonia early.
  • the sensitivity and specificity of the test results of the kit for diagnosing pneumonia reached 99% and 96%, respectively, and it is an excellent test kit.
  • This kit for screening for novel coronavirus infection includes:
  • SARS-CoV-2 magnetic bead coating (R1): Tris buffer containing magnetic bead coating of SARS-CoV-2 recombinant antigen, pH 7.2, containing 0.1% spit Temperature 20 (v/v), 5% bovine serum albumin (w/v), 0.5% Proclin300 (v/v); magnetic beads are used as the reaction solid phase carrier, and the antigen is used to bind the target antibody in the test object.
  • Anti-human IgA antibody acridinium ester marker (R2) 2-(N-morpholine) ethanesulfonic acid buffer solution of acridinium ester-labeled mouse anti-human IgA monoclonal antibody, containing 0.1% Tween 20 (v /v), 5% bovine serum albumin (w/v), 0.5% Proclin300 (v/v). Combining with the antigen-antibody complex, under alkaline conditions, the acridinium ester can emit light after being excited.
  • Reaction buffer (R3) phosphate buffer containing goat anti-human IgG antibody, containing 0.1% Tween 20 (v/v), 5% bovine serum albumin (w/v), 0.5% Proclin 300 ( v/v). Used to adsorb IgG in the sample to be tested to prevent interference.
  • SARS-CoV-2 IgA negative control SARS-CoV-2 IgA negative serum.
  • SARS-CoV-2 IgA positive control It is a human-derived purified SARS-CoV-2 IgM antibody.
  • Negative and positive control information card contains the luminescence value of the negative and positive control, which is used for correction during screening.
  • the experiment also requires the substrate solution (aqueous solution containing NaOH) and the cleaning solution (phosphate buffer containing surfactant) as general reagents, which are not provided in this kit.
  • SARS-CoV-2 IgA turns positive earlier than IgM antibodies, 1-4 days earlier on average. Therefore, the clinical detection of SARS-CoV-2 IgA antibodies can be carried out earlier, and some patients with new coronary pneumonia can be screened and dealt with early.
  • the RBD protein was used as a magnetic bead-coated antigen kit for detection, and clinical studies were conducted on 174 patients and 202 control populations, and the serum SARS-CoV-2 IgA antibody levels of patients with new coronary pneumonia and the control group were obtained.
  • the results are shown in Figure 1. .
  • the IgA level of patients with new coronary pneumonia is significantly higher than that of the control group.
  • the trend of SARS-CoV-2 IgA and IgM antibodies with the course of the disease is shown in Figure 2.
  • Figure 2 it can be seen that the patient’s SARS-CoV-2 IgA turns positive earlier than IgM antibodies, on average 2(1-4) )sky. Therefore, clinical testing of SARS-CoV-2 and IgA antibodies at the time of patient consultation can diagnose some patients with new coronary pneumonia earlier.
  • the N protein was used as a magnetic bead-coated antigen kit for testing.
  • a clinical study was conducted on 100 patients with new coronary pneumonia and 100 controls.
  • the test results showed that the positive rates of SARS-CoV-2 IgA in patients and controls were respectively 98.9% and 4.2%, there is a significant difference between the two groups.
  • the kit described in any one of Examples 1 to 5 was used to detect 10 normal human sera and 10 new coronary pneumonia patients' sera by chemiluminescence immunoassay.
  • the magnetic bead coating (R1) Before installing the machine, the magnetic bead coating (R1) needs to be gently turned upside down about 30 times to make the magnetic bead particles evenly dispersed. There is no need to continue mixing after loading the magnetic bead coating (R1) for the first time.
  • sample application set the sample type as negative control and positive control respectively, and select SARS-CoV-2 IgA project, negative control and The positive control should be made 2 replicates, and click "Run” after confirming.
  • Detection Put the sample in the sample rack (the sample size should be greater than 300 ⁇ L), push it into the sample rack, edit the sample information on the operation interface, select the SARS-CoV-2 IgA project, and click "Run” after confirming.
  • the system will perform the following operations:
  • the total incubation time is 15 minutes.
  • chemiluminescence immunoassay adopts two-step indirect immunoassay.
  • the sample to be tested is incubated with the magnetic bead coating. After magnetic separation and washing of unbound substances, the acridinium ester label is added and incubated together. After washing again, the substrate solution is added, and then the luminescence reaction of the acridinium ester is detected.
  • the substrate solution is added, and then the luminescence reaction of the acridinium ester is detected.
  • the substrate solution is added, and then the luminescence reaction of the acridinium ester is detected.
  • the test result is expressed by the critical value index (COI).
  • the kit of the present invention can detect 100% of patients with new coronary pneumonia, with accurate detection results and high sensitivity.
  • a kit for screening for novel coronavirus infection containing,
  • Test card consists of SARS-CoV-2 IgA test strip and plastic box; the test strip contains nitrocellulose membrane, sample pad, binding pad, absorbent paper, PVC board and other supports; the nitrocellulose membrane is coated with anti-human -IgA antibody and anti-mouse IgG polyclonal antibody, the binding pad contains colloidal gold-labeled SARS-CoV-2 antigen, and the method of obtaining the antigen can be the method in Example 2.
  • Tris hydrochloric acid buffer of the sample diluent The pH of the Tris hydrochloric acid buffer is 7.1-7.4, containing 0.5-2% bovine serum albumin (w/v), 0.05-0.2% Tween 20 (v/v) , 0.25-1% Proclin300 (v/v). Preferably, it contains 1% bovine serum albumin (w/v), 0.1% Tween 20 (v/v), and 0.5% Proclin 300 (v/v).
  • the detection principle of the colloidal gold method is to use the principle of immunochromatographic technology to qualitatively detect SARS-CoV-2 IgA in human serum, plasma, and whole blood.
  • the sample contains SARS-CoV-2 IgA
  • it is combined with the SARS-CoV-2 CoV-2 antigen binds to form a reaction complex.
  • the complex moves forward along the nitrocellulose membrane, and is captured by the anti-human-IgA antibody pre-coated on the detection zone (T) of the nitrocellulose membrane.
  • the detection area (T) agglutinates to form a red reaction line visible to the naked eye.
  • the result is positive; when the SARS-CoV-2 IgA in the sample is below the minimum detection limit, no red reaction line appears in the detection area (T). The result was negative.
  • the kit of Example 1 was used to detect 10 normal human sera and 10 new coronary pneumonia patients with the colloidal gold method.
  • Room temperature refers to a temperature of 10°C to 30°C and a humidity of 45% to 75%.
  • the kit of the present invention can detect 100% of patients with new coronary pneumonia, with accurate detection results and high sensitivity.
  • the present invention also provides the use of a reagent for detecting SARS-CoV-2 IgA antibody in the preparation of the above-mentioned kit for screening new coronavirus infections and the preparation of the reagent for detecting SARS-CoV-2 IgA antibody for screening novel coronavirus infections.
  • a reagent for detecting SARS-CoV-2 IgA antibody in the preparation of the above-mentioned kit for screening new coronavirus infections and the preparation of the reagent for detecting SARS-CoV-2 IgA antibody for screening novel coronavirus infections.
  • Use of coronavirus infection reagents can detect new coronaviruses, can be used to supplement the diagnosis of new coronary pneumonia at an earlier stage, and has the characteristics of accurate detection results.

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Abstract

L'invention concerne un kit de criblage de nouvelles infections par le coronavirus et un réactif de détection associé. Le kit contient un matériau de revêtement de billes magnétiques de SARS-CoV-2 servant de R1 et un marqueur ester d'acridinium d'anticorps anti-IgA humaines servant de R2, le matériau de revêtement de billes magnétiques de SARS-CoV-2 contenant une solution tampon de trihydroxyméthyle aminométhane de matériau de revêtement de billes magnétiques (avec une valeur de pH de 7,1 à 7,4) revêtue d'un antigène recombinant du SARS-CoV-2, et le marqueur ester d'acridinium d'anticorps anti-IgA humaines est une solution tampon d'acide 2-(N-morpholino)éthanesulfonique contenant un anticorps monoclonal de souris anti-IgA humaines marqué par un ester d'acridinium.
PCT/CN2020/080524 2020-03-20 2020-03-20 Procédé de criblage précoce du nouveau coronavirus Ceased WO2021184391A1 (fr)

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115109167A (zh) * 2022-06-30 2022-09-27 宁波博肽生物技术有限公司 一种新型冠状病毒（SARS-CoV-2）重组抗原及其应用
CN116519955A (zh) * 2023-04-11 2023-08-01 四川沃文特生物技术有限公司 用于测定n末端脑利钠肽前体的缓冲液、体系及试剂盒
CN116593696A (zh) * 2023-02-24 2023-08-15 杭州迈尔德生物科技有限公司 一种口含式唾液抗新型冠状病毒S蛋白分泌型IgA抗体检测试剂盒
US11740240B2 (en) 2020-07-20 2023-08-29 Bio-Rad Laboratories, Inc. Immunoassay for SARS-CoV-2 neutralizing antibodies and materials therefor
TWI816315B (zh) * 2022-03-02 2023-09-21 慈濟學校財團法人慈濟大學 表現新型冠狀病毒sars-cov-2蛋白質的大腸桿菌

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