CN111781349A - Diagnostic kit capable of predicting prognosis of COVID-19 patient - Google Patents
Diagnostic kit capable of predicting prognosis of COVID-19 patient Download PDFInfo
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- CN111781349A CN111781349A CN202010394047.6A CN202010394047A CN111781349A CN 111781349 A CN111781349 A CN 111781349A CN 202010394047 A CN202010394047 A CN 202010394047A CN 111781349 A CN111781349 A CN 111781349A
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- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
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- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54306—Solid-phase reaction mechanisms
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- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic
- G01N33/545—Synthetic resin
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/165—Coronaviridae, e.g. avian infectious bronchitis virus
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2469/00—Immunoassays for the detection of microorganisms
- G01N2469/20—Detection of antibodies in sample from host which are directed against antigens from microorganisms
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
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Abstract
The present invention relates to diagnostic kits capable of predicting prognosis of a patient with COVID-19. The novel coronavirus SARS-CoV-2 antibody (IgG) detection device comprises a solid carrier and a probe connected with the solid carrier and having the sequence shown in SEQ ID NO: 1-2.
Description
Technical Field
The invention mainly relates to an antibody detection kit. In particular, the present invention relates to a novel coronavirus (SARS-CoV-2) IgG antibody detection kit, which can be used as a diagnostic kit for predicting prognosis of patients with novel coronavirus pneumonia (COVID-19).
Background
Aiming at the pneumonia (COVID-19) epidemic situation caused by the novel coronavirus (SARS-CoV-2), the diagnostic kit becomes an indispensable tool for preventing and controlling the epidemic situation.
COVID-19 is an acute respiratory infectious disease caused by SARS-CoV-2 virus. Although mortality rates were lower than those of SARS and MERS, the prevalence of SARS-CoV-2 was demonstrated to be much higher, and according to clinical classification, COVID-19 patients were classified into mild, moderate, severe and severe types, with disease severity positively correlated with viral load. Thus, preventing mild to severe progression in patients is likely to be the most effective method for treating SARS-CoV-2 infection.
Therefore, there is an urgent need to develop a diagnostic kit capable of predicting the prognosis of patients with novel coronavirus pneumonia (COVID-19).
Disclosure of Invention
In view of the problems in the prior art, the inventors developed a "short peptide protein composite chip" based on "iPDMS nanomembrane" and "multi-phase antibody kinetics", thereby solving the above technical problems.
Namely, the present invention comprises:
1. a novel coronavirus SARS-CoV-2 antibody (IgG) detection device (detection device 1 of the present invention) comprising a solid support, and the following polypeptide combinations attached to the solid support:
seq ID NO: 1 (S44, GCVIAWNSNNLDSKVGGNYN), and
seq ID NO: 2 (S39, GVSPTKLNDLCFTNVYADSF).
The detection device 1 may be substituted with SEQ ID NO: 1-2, and does not comprise other polypeptides.
2. The detection device of claim 1, wherein the solid support is polydimethylsiloxane (iPDMS membrane) with an initiator on the surface. The polydimethylsiloxane having an initiator on the surface can be found in, for example, chinese patent application publication CN 101265329A.
3. A novel coronavirus SARS-CoV-2 antibody (IgG) detection kit (detection kit 1 of the present invention) comprising the detection device of claim 1 or 2.
4. The test kit according to claim 3, which is used for predicting whether or not a patient suffering from pneumonia caused by a novel coronavirus SARS-CoV-2 of an ordinary type or a severe type will have an improved disease state.
Here, the improvement of the disease includes the transformation of nucleic acid detection to negative, the alleviation of clinical symptoms, the transformation of severe form to general form or mild form, the transformation of general form to mild form, and the compliance with clinical rehabilitation standards.
5. Use of the detector device of claim 1 or 2 for the preparation of a novel coronavirus SARS-CoV-2 antibody (IgG) detection kit for predicting whether a patient with pneumonia caused by a novel coronavirus SARS-CoV-2 of general or severe type will have an improved disease.
The detection device 1 or the detection kit 1 of the present invention can be used for predicting whether or not the patient's condition of pneumonia caused by the novel coronavirus SARS-CoV-2 of normal type or severe type will be improved. In particular to the method for preparing the nano-particles,
for a patient clinically determined to be of the general type or the severe type, blood is collected at 1 to 3 days/time and tested by the test kit 1, and 2 or more, preferably 3 or more, more preferably 4 or more, more preferably 5 or more, more preferably 10 or more determinations are made continuously or at intervals, and for any one of the above polypeptides, if there is a response in 3 or more, preferably 4 or more, more preferably 5 or more determinations continuously, it is determined that the patient's condition will be improved; otherwise, the patient's condition is judged not to improve.
In this specification, "response" means that the signal value of a positive spot read with the reading device is significantly different from the signal value of a negative spot. For example, the signal value of a positive spot read by the reading device is greater than or equal to 10, preferably greater than or equal to 20, more preferably greater than or equal to 30; and the signal value of the negative spot read by the reading device is less than 10, preferably less than 5, more preferably less than 1. The recognition device may be, for example, a microarray chip imager manufactured by abbotto biotechnology, su.
In the present specification, the solid support may be one or a plurality of solid supports, but preferably one, that is, all the polypeptides are independently attached to the same solid support. In the present invention, the solid carrier is not particularly limited as long as it is a carrier which is a solid or an insoluble material. The polypeptide can be linked to the solid support by methods known to those skilled in the art.
Examples
Example 1 preparation and validation of Polypeptides and proteins
SEQ ID NO: 1-2 by Nanjing Kinsrui Biotechnology Ltd.
Example 2 preparation of a kit (detection device)
Kit (detection device) 1
Respectively spotting the SEQ ID NO: 1-2, and spotting positive and negative quality control spots to prepare a kit (detection device) 1 (48-well plate type chip reaction plate).
Example 3 detection Using the kit
Inspection step
1. Preparation work: the reagent and the test sample need to be balanced to room temperature before the test; serum was diluted 100-fold with serum diluent (50-fold dilution of whole blood).
2. Chip wetting: the chip reaction plate is taken out. The chip surface was soaked with the wash solution for 3 minutes and then the wash solution was discarded.
3. Adding a sample: 100 mul of sample to be tested is added into the reaction hole of the chip.
4. Sample incubation: the chip reaction plate was placed in a constant temperature incubator and incubated at 37 ℃ and 500rpm for 30 minutes.
5. Cleaning a sample: the sample in the reaction well of the chip is discarded, and the reaction well is rinsed with washing solution and spun off, and repeated 3 times.
6. Adding enzyme antibody: and adding 100 mu L of enzyme-labeled antibody liquid into the reaction hole of the chip, wherein the enzyme-labeled antibody is a goat anti-human IgG antibody marked by HRP.
7. And (3) enzyme-resistant incubation: the chip reaction plate was placed in a constant temperature incubator and incubated at 37 ℃ and 500rpm for 30 minutes.
8. Enzyme anti-cleaning: and (3) discarding the enzyme-labeled antibody liquid in the reaction hole of the chip, washing the reaction hole by using a cleaning solution, throwing away the reaction hole, and repeating for 3 times.
9. Color development: adding 70 μ L of chromogenic substrate solution into each hole, and standing at room temperature for 5-10 min.
10. Discarding the color developing solution, prying the cover (using a straight screwdriver), rinsing with ultrapure water for 3 times, and drying in the air/blow-drying.
11. The developed signal was automatically collected by photographing with a 48-well microarray chip imager (manufactured by Suzhou Albout Biotechnology Co., Ltd.).
12. And (4) judging a result:
for 7 patients (Severe 5, general 2) with COVID-19, blood was collected for 3 days/time and tested by the kit 1, and the test was continued for 5 or more times, and if any of the polypeptides in the kit 1 had a response (signal value measured by the instrument was greater than 10) in 3 or more times of the tests, the patients were predicted to have an improvement (positive); otherwise, the patient is predicted to have no improvement (negative).
For 5 of the 7 COVID-19 patients (Severe 4, Normal 1), the patient was predicted to have an improvement; in fact, the patient's condition improved within 7 days from the time point of the first measurement of the above-mentioned 3 or more consecutive measurements (including negative conversion from nucleic acid detection, ICU removal, disappearance of hyperthermia, conversion from severe to normal or mild, and conversion from normal to mild).
For 2 of the 7 COVID-19 patients (Severe 1, general 1), no improvement was predicted; in fact, during the blood collection and measurement period, the condition of the patient is not improved or even worsened.
Therefore, the kit of the present invention can be used for evaluating the improvement of the patients with pneumonia caused by the novel coronavirus SARS-CoV-2 of normal type or severe type.
While embodiments of the present invention have been described above, the present invention is not limited to the specific embodiments and fields of application described above, which are intended to be illustrative, instructive, and not limiting. Those skilled in the art, having the benefit of this disclosure, may effect numerous modifications thereto without departing from the scope of the invention as defined by the appended claims.
Claims (5)
1. A novel coronavirus SARS-CoV-2 antibody (IgG) detection device, comprising a solid support, and the following polypeptide combinations linked to the solid support:
seq ID NO: 1 (S44, GCVIAWNSNNLDSKVGGNYN), and
seq ID NO: 2 (S39, GVSPTKLNDLCFTNVYADSF).
2. The detection device of claim 1, wherein the solid support is polydimethylsiloxane having an initiator on its surface.
3. A novel coronavirus SARS-CoV-2 antibody (IgG) detection kit comprising the detection device of claim 1 or 2.
4. The test kit according to claim 3, which is used for predicting whether or not a patient suffering from pneumonia caused by a novel coronavirus SARS-CoV-2 of an ordinary type or a severe type will have an improved disease state.
5. Use of the detector device of claim 1 or 2 for the preparation of a novel coronavirus SARS-CoV-2 antibody (IgG) detection kit for predicting whether a patient with pneumonia caused by a novel coronavirus SARS-CoV-2 of general or severe type will have an improved disease.
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CN2020102609965 | 2020-04-03 | ||
CN202010260996 | 2020-04-03 |
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CN111781349A true CN111781349A (en) | 2020-10-16 |
CN111781349B CN111781349B (en) | 2022-04-22 |
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1570638A (en) * | 2003-07-22 | 2005-01-26 | 李克生 | SARS virus antibody detecting method, rapid diagnosis kit and preparation method |
US20050112559A1 (en) * | 2003-09-29 | 2005-05-26 | The Chinese University Of Hong Kong | Compositions and methods for diagnosing and preventing severe acute respiratory syndrome (SARS) |
CN104945476A (en) * | 2014-12-31 | 2015-09-30 | 苏州偲聚生物材料有限公司 | Polypeptide, detection device comprising polypeptide, and detection kit comprising device |
WO2016090345A1 (en) * | 2014-12-05 | 2016-06-09 | Planet Biotechnology Inc. | Dpp4 immunoadhesin compositions and methods |
Family Cites Families (4)
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CN105785026B (en) * | 2014-12-24 | 2017-07-18 | 中国科学院苏州纳米技术与纳米仿生研究所 | Kit and detection method for detecting enterovirns type 71 IgM antibody |
CN104945477A (en) * | 2014-12-31 | 2015-09-30 | 苏州九龙医院有限公司 | Polypeptide, detection device comprising polypeptide, and detection kit comprising device |
CN107090024A (en) * | 2016-02-18 | 2017-08-25 | 中国科学院苏州纳米技术与纳米仿生研究所 | Detection device, detection kit and diagnostic method available for diagnostic system lupus erythematosus |
EP3530668A1 (en) * | 2018-02-22 | 2019-08-28 | Euroimmun Medizinische Labordiagnostika AG | A novel assay for the diagnosis of viral infections |
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- 2020-05-11 CN CN202010394047.6A patent/CN111781349B/en active Active
- 2020-05-25 CN CN202010446995.XA patent/CN111679084B/en active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1570638A (en) * | 2003-07-22 | 2005-01-26 | 李克生 | SARS virus antibody detecting method, rapid diagnosis kit and preparation method |
US20050112559A1 (en) * | 2003-09-29 | 2005-05-26 | The Chinese University Of Hong Kong | Compositions and methods for diagnosing and preventing severe acute respiratory syndrome (SARS) |
WO2016090345A1 (en) * | 2014-12-05 | 2016-06-09 | Planet Biotechnology Inc. | Dpp4 immunoadhesin compositions and methods |
CN104945476A (en) * | 2014-12-31 | 2015-09-30 | 苏州偲聚生物材料有限公司 | Polypeptide, detection device comprising polypeptide, and detection kit comprising device |
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CN111679084A (en) | 2020-09-18 |
CN111781349B (en) | 2022-04-22 |
CN111679084B (en) | 2022-07-05 |
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