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Method for diagnosing SARS-CoV-2 infection

A sars-cov-2, protein technology, applied in the field of SARS-CoV-2 infection diagnosis, can solve the problems of invasion, inconvenient sampling, poor diagnostic accuracy, etc., achieve high accuracy, save medical care costs, facilitate sampling and quick effect

Pending Publication Date: 2021-11-09
UNIV OF SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is inconvenient to take samples, is not easy to accept, requires the participation of professional medical staff, and has the risk of infectious diseases; in addition, because the viral load in the pharynx will decrease significantly with the increase of infection time, especially after 15 days of infection, Prone to false negative test results, poor diagnostic accuracy
[0003] In addition to the above RT-qPCR detection methods that are in general use, methods for diagnosing SARS-CoV-2 infection by detecting SARS-CoV-2-specific antibodies in serum, such as IgM and IgG, are also emerging, but blood collection is a Intrusive behavior, and requires the participation of professional nursing staff, it is difficult to apply to places that require rapid and large-scale detection, such as airports and customs

Method used

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  • Method for diagnosing SARS-CoV-2 infection
  • Method for diagnosing SARS-CoV-2 infection
  • Method for diagnosing SARS-CoV-2 infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Collection and treatment of saliva samples

[0031] 1) Randomly select two healthy people, as well as four COVID-19 rehabilitation patients as the detected detectors 1, 2, 3, 4, 5, 6, and 6. Inform the detected person, it is not possible to perform diet, mouthwash, etc., can affect the saliva quality in at least 10 minutes before taking a sample. At the same time, the saliva collection container is issued to the detected, and the collection container opening is preferably greater than 5 cm and has a sealing cover.

[0032] 2) Clean the surface of the container on the surface of about 1 ml of saliva.

[0033] 3) Collect the sampled saliva samples of the detected, and use 75% alcohol to disinfect the outer surface of the vessel.

[0034] 4) -20 ° C Save, open in the biosafety cabinet during detection.

Embodiment 2

[0035] Example 2. SARS-COV-2 specific IgA in saliva samples using immunopolymerization method

[0036] 1) Expression of the recombinant binding region of purification of SARS-COV-2 ugly proteins (SEQUENCE ID: MT322424.1, specific nucleic acid sequence SEQ ID NO.1 See the last page of the specification), and coupled to CNBR-ACTIVATED Sepharose TM 4B Agarose beads (purchased from GE).

[0037] 2) Take the above 2 healthy people and 4 COVID-19 rehabilitation patients (i.e., detected 1, 2, 3, 4, 5, 6) each 1 ml of each of which were diluted with 4 ml of PBS and transferred to 10 ml of centrifuge tube, respectively. Each addition of 100 μL of the agarose beads of the coupling RBD prepared in the first embodiment of the present embodiment, mixing, and incubated at room temperature for 30 min.

[0038] 3) 1000g Centrifuge 1min and discard it. Then, 4 ml of PBS was added, mixed upside down 20 times to clean the beads.

[0039] 4) 1000g Centrifuge 1min, repeating this Example 2 Step 3) 4 t...

Embodiment 3

[0047] Example 3. The detection principle of SARS-COV-2 specific IgA utilization chemiluminescence method was detected by chemiluminescence method: the sample to be tested by saliva and the magnetic beads with SARS-COV-2 RBD After incubation, after the washing washing washed, the anti-human IgA antibody acridate syrids were added to incubate together, and after washing, the substrate fluid was added, and the luminescence reaction of the acridine ester was then detected. If there is a new coronavirus IgA antibody in the sample, a magnetic bead package can be formed to form a positive correlation between the luminescence intensity of the acridine ester with a new coronavirus IgA antibody. Relationship, the test results are expressed in the critical value index (COI).

[0048] The chemiluminescent detection of this embodiment is used in full automatic detection machine Kreser 1000, a yin-positive control, and is used to screen the correction.

[0049] The specific steps are:

[0050]...

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Abstract

The invention discloses a method for diagnosing SARS-CoV-2 infection by detecting SARS-CoV-2 specific IgA (Immunoglobulin A) in saliva. The diagnosis method can be any method capable of detecting IgA, such as ELISA (Enzyme-Linked Immunosorbent Assay), co-immunoprecipitation, chemiluminescence and a colloidal gold method. The method proves that there is SARS-CoV-2 specific IgA in saliva of a COVID-19 patient, and can be used for clinical diagnosis of SARS-CoV-2 infection.

Description

Technical field [0001] The invention belongs to the field of antibody detection, and more particularly to methods of SARS-COV-2 infection diagnosis by detecting IgA anti-SARS-COV-2 in saliva. Background technique [0002] Currently, the accurate diagnosis and isolation caused by SARS-COV-2 or 2019-NCOV remains the main method of controlling the epidemic. The main method of detecting the diagnosis of SARS-COV-2 infections worldwide is the use of reverse transcription real-time quantitative PCR (RT-QPCR) to detect the viral RNA on the nasopharyngeal swab. This method is not convenient to be sampled, not easy to accept, require professional medical staff to participate, and risk of infectious diseases; It is prone to false negative test results, and the diagnostic accuracy is poor. [0003] Except for the general RT-QPCR detection method, the method of detecting SARS-COV-2 specific antibodies in serum, such as IgM and IgG, to diagnose SARS-COV-2 infection, but blood is a kind Invasi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/569
CPCG01N33/68G01N33/56983G01N2333/165G01N33/6854G01N2469/20
Inventor 金腾川马欢曾威红
Owner UNIV OF SCI & TECH OF CHINA
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