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Novel coronavirus neutralizing antibody detection kit based on magnetic particle chemiluminescence and application thereof

A chemiluminescence and antibody detection technology, applied in the field of clinical testing, to achieve the effect of easy operation, low cost and high stability

Pending Publication Date: 2021-06-22
山东莱博生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to the principle that the luminescence value is proportional to the effective concentration of the RBD neutralizing antibody in the sample, the preparation of the new coronavirus neutralizing antibody detection kit based on magnetic particle chemiluminescence and its application have not been reported yet.

Method used

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  • Novel coronavirus neutralizing antibody detection kit based on magnetic particle chemiluminescence and application thereof
  • Novel coronavirus neutralizing antibody detection kit based on magnetic particle chemiluminescence and application thereof
  • Novel coronavirus neutralizing antibody detection kit based on magnetic particle chemiluminescence and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1: Preparation of a neogular virus and antibody detection tester according to a magnetic particulate chemiluminescence of the present invention

[0073] (1) Preparation of magnetic microspheres:

[0074] 1.1) Formulate the coupling buffer: i.e. to formulate 0.01 mol / L MES buffer solution, and adjust the pH of 6.0;

[0075] MES 1.95G

[0076] Purification water to 1000ml

[0077] Filtration and sterilization, stored at 4 ° C;

[0078] 1.2) The formulation and method for preparing the washing liquid, the washing liquid is:

[0079]

[0080] 1.3) Formulation of preservation fluids, preservation fluids is:

[0081]

[0082] 1.4) Coupling operation

[0083] (1) RBD protein 1 acidification treatment

[0084] Acid treatment: Take 1 mL of concentration of 1 mg / ml of RBD protein 1 solution to 2.0 ml of low adsorption EP tube, add 0.2 ml of acidification treatment liquid a to the EP tube, and mix well at room temperature 40 rpm.

[0085] Metallization: 0.4 ml of treat...

Embodiment 2

[0117] Example 2: The use of magneto-particulate chemiluminescent and antibody detection kits based on magneto-particulate chemiluminescence and antibody detection kits in detecting new types of coronary viruses and antibody biological samples.

[0118] Among them, the preferred method of detecting a new type of coronavirus and antibody sample is:

[0119] a) Add 50 μl of sample 5 μl, 50 μl of sample dilution, and add concentration of 0.2-0.5 mg / ml to 50 μl of a concentration of 0.2-0.5 mg / ml to the reaction cup. Positive control, negative control Press a) the same step operation;

[0120] b) incubation for 15 minutes at 37 ° C after mixing;

[0121] c) Wash 4-5 times with magnetic separation frame or magnetic separator.

[0122] d) Add a horseradish peroxidase-labeled recombinant RBD protein 2 solution 100 uL to the fully automatic magnetic particle chemiluminescence analyzer reaction cup;

[0123] e) incubate at 37 ° C for 15 minutes after mixing;

[0124] f) Wash 4-5 times ...

Embodiment 3

[0132] Example 3: Detection of linearity, accuracy, and precision detection of a neogignic micro crown virus and antibody detection kit of a neogignic micro-crown and antibody detection kit

[0133] 1) Linearity of the kit

[0134] (1) The subscript formulation: RBD neutralizing antibody was diluted to 10 ng / ml, 20 ng / ml, 50 ng / ml, 100 ng / ml, 250 ng / ml, 500 ng / ml, 1000 ng / ml.

[0135] (2) Test: The method of formulation is detected by the method described in Example 2 to obtain an RLU value;

[0136] (3) Establishment of standard curve: The calibration curve of the reagent is generated according to the preparation concentration of the calibration and the RLU value employ a suitable fitting manner. See Table 1 and figure 1 .

[0137] Table 1 kit linearity

[0138] concentration 0 ng / ml 10 ng / ml 20 ng / ml 50 ng / ml 100 ng / ml 250ng / ml 500ng / ml 1000 ng / ml RLU 23258 398748 736886 4248972 12038794 40245372 86548971 123549786

...

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Abstract

The invention discloses a novel coronavirus neutralizing antibody detection kit based on magnetic particle chemiluminescence. The novel coronavirus neutralizing antibody detection kit is composed of magnetic microspheres which are coupled with RBD protein 1 and arranged in a kit body, an RBD neutralizing antibody standard substance solution serving as a positive control, a negative control, a sample diluent, an RBD protein 2 enzyme conjugate, a 20*concentrated washing solution and a luminescent solution. The invention also discloses application of the kit in detection of a biological sample containing the novel coronavirus neutralizing antibody. Experiments prove that the kit disclosed by the invention is high in stability, strong in selectivity, high in detection speed, low in cost and easy to operate, overcomes the defects of high laboratory environment requirement, long operation time and the like when the neutralizing antibody is detected in the prior art, and shortens the operation time from 120 minutes to 35 minutes. The kit not only can be used for judging the antibody neutralizing condition after novel coronavirus vaccine inoculation, but also can be used for screening common novel coronavirus antibodies, and has a great clinical application prospect.

Description

Technical field [0001] The present invention relates to a new type of coronavirus (SARS-COV-2) and antibody detection, and more particularly to a new crown virus and antibody detection kit based on magnetic particulate chemiluminescence and its application, which belongs to the technical field of clinical testing. Background technique [0002] New crown virus (SARS-COV-2) is a big population is the most severe challenge for human beings since the century. New coronavirus, "SARS-COV-2" infection, common somewhere with respiratory symptoms, fever, cough, air promotion and difficulty breathing. In a more serious case, infection can lead to pneumonia, severe acute respiratory syndrome, renal failure, and even death. Because of its infectivity, the survival is high, the total number of global infections has been close to 90 million in one year. Experts have generally believed that before developing a safe and effective vaccination and successfully implemented a global vaccination prog...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/58G01N33/569G01N33/543G01N21/76
CPCG01N33/6854G01N33/56983G01N33/54326G01N33/581G01N21/76G01N2469/20G01N2333/165
Inventor 欧兰香陈振王岩丁兴龙武建伟陈文丽
Owner 山东莱博生物科技有限公司
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