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Novel coronavirus (COVID-19) neutralizing antibody detection kit and detection method

A COVID-19 and coronavirus technology, applied in the field of novel coronavirus neutralizing antibody detection kits, can solve problems such as inability to perform quantitative detection, and achieve the effects of improving labeling efficiency, improving analytical sensitivity, and improving detection sensitivity

Pending Publication Date: 2021-07-23
深圳市爱康试剂有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method can only be used for qualitative detection, not quantitative detection, and cannot provide more detection information for clinical practice.

Method used

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  • Novel coronavirus (COVID-19) neutralizing antibody detection kit and detection method
  • Novel coronavirus (COVID-19) neutralizing antibody detection kit and detection method
  • Novel coronavirus (COVID-19) neutralizing antibody detection kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Preparation of reagent cards

[0051] (1) Preparation of test line and quality control line: prepare angiotensin-converting enzyme 2 (ACE2) concentration of 1.0 mg / mL and chicken IgY antibody concentration of 0.5 mg / mL respectively, and use a spray film machine to draw lines onto nitrocellulose In the detection line and quality control line area on the membrane, the coating volume is 1.0 μL / cm, and the nitrocellulose membrane is placed in a forced air drying oven, and dried at 37 ° C for 18 hours;

[0052] (2) Preparation of fluorescent binding pad: the material of the fluorescent binding pad is glass fiber, soaked in the pretreatment solution of the binding pad, dried in a blast drying oven, and then using the conjugate diluent to label the new coronavirus with fluorescent microspheres Spike glycoprotein (RBD) was diluted 20 times, chicken IgY polyclonal antibody labeled with fluorescent microspheres was diluted 50 times, sprayed on the fluorescent binding pad with a s...

Embodiment 2

[0073] Preparation of reagent cards

[0074] (1) Preparation of test line and quality control line: prepare angiotensin-converting enzyme 2 (ACE2) concentration of 2.0 mg / mL and rabbit IgG antibody concentration of 1 mg / mL respectively, and use a spray film machine to streak onto the nitrocellulose membrane In the upper detection line and quality control line area, the coating volume is 2 μL / cm, and the nitrocellulose membrane is placed in a blast drying oven, and dried at 45 ° C for 12 hours;

[0075] (2) Preparation of fluorescent conjugation pads: the material of the fluorescent conjugation pads is polyester film, which is soaked in the conjugation pad pretreatment solution, dried in a blast drying oven, and the novel coronavirus labeled with fluorescent microspheres using conjugate diluent Spike glycoprotein (RBD) was diluted 5 times, chicken IgY polyclonal antibody labeled with fluorescent microspheres was diluted 20 times, sprayed on the fluorescent binding pad with a sp...

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Abstract

The invention discloses a novel coronavirus (COVID-19) neutralizing antibody detection kit, which is characterized by comprising: a detection card. The detection card comprises a card shell and a test strip arranged in the card shell, the test strip comprises a sample pad, a fluorescence combination pad, a nitrocellulose membrane, a water absorption pad and a backing plate. The fluorescent combination pad comprises a detection line fluorescent conjugate and a quality control line fluorescent conjugate, the detection line fluorescent conjugate is a novel recombinant coronavirus spinous process glycoprotein (RBD) marked by fluorescent microspheres, and the nitrocellulose membrane is coated with a detection line formed by angiotensin converting enzyme 2 (ACE2). And the nitrocellulose membrane is also coated with a quality control line formed by IgY, IgG or DNP antibodies. The kit is combined with a fluorescence immunoassay analyzer to detect a sample, and the kit is high in detection sensitivity, good in reproducibility, high in stability and convenient to operate.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a novel coronavirus (COVID-19) neutralizing antibody detection kit and detection method. Background technique [0002] The novel coronavirus (COVID-19) has four main structural proteins: Spike protein (S protein), nucleocapsid protein (Nucleocapsid, N protein), membrane protein (Membrane protein, M protein), envelope Protein (Envelope protein, E protein). The spike protein S protein has two subunits: S1 and S2, and the receptor binding domain (RBD) is located on the S1 subunit. RBD is a unique region of the spike protein antigen, which can bind to the human ACE2 receptor. Through the combination of RBD and ACE-2 receptor, COVID-19 mediates the virus to enter the interior of the cell and reproduce, resulting in symptoms and leading to disease. [0003] The neutralizing antibody of the new coronavirus can specifically recognize and bind the RBD site of the new coronavirus, bind to th...

Claims

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Application Information

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IPC IPC(8): G01N33/58G01N33/68G01N33/569G01N33/544G01N33/558G01N21/64
CPCG01N21/6428G01N33/544G01N33/558G01N33/56983G01N33/582G01N33/585G01N33/6854G01N2333/165
Inventor 陈雪肖宇强陈粤秀张平
Owner 深圳市爱康试剂有限公司
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