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Novel coronavirus detection kit

A detection kit and coronavirus technology, which is applied in the field of colloidal gold kits and its preparation for novel coronaviruses, can solve the problems of missed detection of detection results, missed detection, N protein cannot accurately label antibodies, etc., and achieve simple sampling , avoid false negatives, and prevent the spread of the epidemic

Pending Publication Date: 2020-06-26
SHANXI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, in the early stage of 2019-nCoV virus infection, due to the extremely small amount of antibodies produced in the body, the N protein may not be able to accurately label the antibodies in the body, which may lead to missed detection
[0010] At the same time, the detection sensitivity of the kit is usually lower than that of nucleic acid detection, which will also cause missed detection in the detection results.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] 1) Preparation of colloidal gold solution: add 500mL ultrapure water to a round bottom flask, then add 10mL of 1% gold chloride solution, heat with stirring until it just starts to boil, then quickly add 12mL of 10% trisodium citrate solution at one time, After the solution turned deep red completely, the heating was stopped, and the reaction was continued for 10 minutes using residual heat. Then cool down to room temperature with cold water, add deionized water to make up to 500mL, and store at 4°C in the dark.

[0050] 2) Antigen labeling: take 10mL colloidal gold solution, add 60μL 0.1mol / L potassium carbonate solution, quickly add 100μg N protein antigen, mix well and place at room temperature for 30 minutes. Quickly add 100 μL of 20% bovine serum albumin solution, mix well, place at room temperature for 30 minutes, and centrifuge at 12,000 rpm for 20 minutes. Discard the supernatant, then add 5 mL of 50 mmol / L, pH7.4 phosphate buffer to suspend the precipitate, ce...

Embodiment 2

[0058] Example 2: Use of the novel coronavirus detection kit.

[0059] 1) Preparation: First, put the kit and blood sample at room temperature to return to normal temperature.

[0060] 2) Tear open along the incision of the packaging bag, take out the kit and put it on the horizontal platform and press the sample number.

[0061] 3) Add 10 μL of plasma or serum to the sample well, and then immediately add 80 μL of sample diluent.

[0062] 4) After 10 minutes, read the result in the result observation area and make a judgment.

[0063] Judgment criteria such as figure 2 .

[0064] Positive: C zone (quality control line) shows a red line, T zone (test line) shows one or two red lines ( figure 2 A, B, C).

[0065] Negative: C area (quality control line) shows a red line, T area (test line) has no color ( figure 2 D).

[0066] Invalid: Area C (quality control line) has no color development ( figure 2 E, F, G, H).

Embodiment 3

[0067] Example 3: Comparison of a single antigen kit and two antigen kits.

[0068] According to the method of Example 1, a kit A in which only the N protein antigen is coated on the colloidal gold pad, and a kit B in which the antigens are N & S-RBD two proteins are prepared respectively.

[0069] The serum / plasma of 40 patients with clinically confirmed COVID-19 from the First Hospital of Shanxi Medical University and the Affiliated Pulmonary Hospital of Shanxi Medical University and the serum / plasma of 30 healthy people were obtained, and tested with A and B kits respectively. The test results are shown in Table 1.

[0070]

[0071] It can be seen from the results in Table 1 that the detection sensitivity of kit B using the two proteins of N & S-RBD is significantly higher than that of kit A, indicating that adding S-RBD protein as an antigen can reduce the rate of false detection and missed detection, and improve the initial detection rate. screening accuracy.

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Abstract

The invention discloses a novel coronavirus detection kit. The kit comprises a bottom plate, and a sample pad, a colloidal gold pad, an NC membrane and a water absorption pad which are fixed on the bottom plate, wherein a detection line and a quality control line are arranged on the NC membrane, an antigen coated on the colloidal gold pad is a mixed antigen protein composed of N protein marked bycolloidal gold and S-RBD protein marked by colloidal gold, and the detection line is a mouse anti-human IgM antibody and mouse anti-human IgG antibody detection line. The detection kit provided by theinvention takes N protein and S-RBD protein as antigens, can improve the sensitivity of COVID-19 preliminary detection, and is suitable for quick screening of large-scale crowds.

Description

technical field [0001] The invention relates to a colloidal gold detection kit, in particular to a colloidal gold detection kit for novel coronavirus and a preparation method thereof. Background technique [0002] The 2019 novel coronavirus (2019-nCoV) is currently the seventh known coronavirus that can infect humans after SARS and MERS. 2019-nCoV is mainly transmitted through respiratory droplets and contact, and has the characteristics of long incubation period, strong infectivity, and asymptomatic infection. Therefore, the development of effective diagnostic kits is the key technical support for the effective prevention and control of the new crown pneumonia epidemic. [0003] The current laboratory detection method for 2019-nCoV infection is mainly viral nucleic acid detection, which is the gold standard for clinical diagnosis of suspected cases. However, this detection method is cumbersome, costly, and has a long detection cycle, and it needs to be carried out in a ce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/58G01N33/577G01N33/569G01N33/558
CPCG01N33/6854G01N33/587G01N33/577G01N33/56983G01N33/558G01N2333/165G01N2469/20
Inventor 解军王磊王晓玲侯淑琳刘志贞黄婷娟李秀娟
Owner SHANXI MEDICAL UNIV
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