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Novel coronavirus IgA antibody magnetic particle chemiluminiscence method detection kit

A chemiluminescence method and detection kit technology, which can be used in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, measurement devices, etc., which can solve the problem that the detection of new coronavirus-specific IgA antibodies is not popularized.

Pending Publication Date: 2020-12-18
BIOSCIENCE (CHONGQING) DIAGNOSTIC TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, the diagnostic products for the new coronavirus at home and abroad are mainly total antibody and combined detection reagents for IgM and IgG antibodies, while the detection of the new coronavirus-specific IgA antibody has not yet been popularized.

Method used

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  • Novel coronavirus IgA antibody magnetic particle chemiluminiscence method detection kit
  • Novel coronavirus IgA antibody magnetic particle chemiluminiscence method detection kit
  • Novel coronavirus IgA antibody magnetic particle chemiluminiscence method detection kit

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preparation example Construction

[0058] The preparation of the magnetic particle-anti-FITC antibody includes the following steps: activate the ferric oxide microspheres with glutaraldehyde, mix well at room temperature for 6 hours, wash three times with 0.05M PBS buffer solution with pH=7.4, and use the Suspend the solution with a concentration of 50mg / mL. After that, add 100ug of mouse anti-FITC antibody to each milliliter suspension, mix and incubate at 37°C for 4h, buffer with an equal volume of 0.05M PBS mass fraction of 5% BSA pH=7.4 The liquid was blocked at 37° C. for 1 hour; finally, washed three times with 0.5% BSA 0.02M Tris-HCl pH=8.0 buffer solution; the particle size of the ferric oxide microspheres was 0.2 μm.

[0059] Preparation of new coronavirus dilution solution: Dissolve 2.1g BSA in 2.1L 0.05M PBS buffer solution, the pH range is 7.5, stir until completely dissolved, and add 2.1mL ProClin TM Stir and mix at 300 for 30min.

[0060] The preparation of FITC-labeled novel coronavirus recombin...

Embodiment 1

[0073] The preparation of horseradish peroxidase-labeled mouse anti-human IgA monoclonal antibody comprises the steps:

[0074] A: Horseradish peroxidase (HRP) activation

[0075] 1) Prepare 10mg / mL HRP solution;

[0076] 2) Prepare 12.8mg / mL sodium periodate NaIO 4 solution;

[0077] 3) Mix the solution prepared in step 1) and step 2) at a volume ratio of 2:1, and react at 4°C in the dark for 30 minutes;

[0078] 4) Prepare an aqueous ethylene glycol solution with a concentration of 20 μL / mL, mix it with the solution prepared in step 3) in the same volume, react at room temperature in the dark for 30 minutes, and the activation is complete, and store at -20°C (the storage time does not exceed 3 months) ;

[0079] B. Horseradish peroxidase-labeled mouse anti-human IgM monoclonal antibody

[0080] 1) Place the IgA monoclonal antibody to be labeled in a dialysis bag, the dialysis buffer is 0.02M carbonate buffer, and dialyze for 1 hour.

[0081] 2) Mix the above-dialyzed m...

Embodiment 2

[0121] The preparation of alkaline phosphatase-labeled mouse anti-human IgA monoclonal antibody comprises the steps:

[0122] 1) Place the IgA monoclonal antibody to be labeled in a dialysis bag, the dialysis buffer is 0.02M carbonate buffer, and dialyze for 1 hour.

[0123] 2) Mix the above dialyzed monoclonal antibody and activated alkaline phosphatase at a mass ratio of 1:0.8, and then dialyze in 0.02M carbonate buffer for 8 hours.

[0124] 3) Prepare NaBH with a concentration of 2mg / mL 4 Aqueous solution, according to 1mgALP plus 80μL prepared NaBH 4 The ratio of the aqueous solution was mixed, and reacted at 4°C in the dark for 2 hours;

[0125] 4) Dialyze the labeling solution completed in step 3) with 0.01M PBS at 4°C for 24 hours, add an equal volume of glycerol, and store at -20°C.

[0126] Chemiluminescent substrates were prepared as follows:

[0127] 1) Measure 900mL of purified water;

[0128] 2) Add 0.2gAMPPD, 0.05g Na respectively to the purified water of st...

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Abstract

The invention provides a novel coronavirus IgA antibody magnetic particle chemiluminiscence method detection kit. The novel coronavirus IgA antibody magnetic particle method detection kit comprises amagnetic particle anti-FITC antibody, an FITC labeled novel coronavirus recombinant antigen and an alkaline phosphatase or horseradish peroxidase labeled human IgA monoclonal antibody. The kit provided by the invention can well supplement diagnosis of a nucleic acid negative new crown infection patient, also shows a good value for diagnosis of an asymptomatic infection patient, can realize early diagnosis and early treatment, reduces a window period as much as possible, and has an important research value for early diagnosis of suspected cases.

Description

technical field [0001] The invention belongs to the technical field of immunoassay and detection, and in particular relates to a novel coronavirus IgA antibody magnetic particle chemiluminescence detection kit. Background technique [0002] The viral load is high during the first infection and repeated exposure to the virus, and specific proteins of the pathogen, such as the S protein and N protein of Covid-19, will stimulate the immune system to cause an antibody response. [0003] T.M.Gallagher's research found that the coronavirus spike protein (S protein) is a type I transmembrane envelope glycoprotein, which is composed of two domains, S1 and S2, which are responsible for binding and fusion with the virus. The middle fragment of S1 is a receptor The body-binding domain can specifically bind to the angiotensin-converting enzyme (ACE2) on the target cell, protrude from the surface of the virus, and can contact the host cell earlier, and is the main inducer of neutralizing...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/577G01N33/543G01N33/535G01N21/76
CPCG01N33/56983G01N33/577G01N33/535G01N33/54326G01N21/76G01N2333/165G01N2469/20
Inventor 刘萍栾大伟张振斌史光辉何景珍田鸽
Owner BIOSCIENCE (CHONGQING) DIAGNOSTIC TECH CO LTD
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