Coxsackie virus A16 (CA16) detection testing strip

A Coxsackie virus and detection test paper technology, applied to measuring devices, instruments, scientific instruments, etc., to achieve the effects of simple operation, clear and easy to distinguish results, and easy promotion

Inactive Publication Date: 2012-02-15
BEIJING ZHUANGDI HAOHE BIOMEDICINE SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, so far, there is no report on a detection s

Method used

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  • Coxsackie virus A16 (CA16) detection testing strip
  • Coxsackie virus A16 (CA16) detection testing strip
  • Coxsackie virus A16 (CA16) detection testing strip

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1: Preparation of anti-Coxsackie virus A16 type VP1 protein monoclonal antibody

[0030] (1) Immunized mice

[0031] After the prepared genetically engineered antigen was taken out from the -20 low-temperature refrigerator and dissolved, it was subcutaneously injected into the back of BALB / C mice (0.2 mg / mouse) with an interval of 10 days. Three days before fusion, mice were challenged with 0.1 mg of antigen in the peritoneal cavity. The immune effect was detected by ELISA method.

[0032] (2) Myeloma cells

[0033] SP2 / 0 myeloma cells: purchased from the Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences. Resuscitate the SP2 / 0 cells stored in the liquid nitrogen tank, and culture them in DMEM medium containing 10% calf serum for 48-72 hours. Number split, ready to fuse.

[0034] (3) cell fusion

[0035] Prepared SP2 / 0 cells and splenic lymphocytes of immunized BALB / C mice were prepared in 2×10 7 / ml and 1×10 8 / ml. Take...

Embodiment 2

[0042] Embodiment 2: Assay of anti-Coxsackie virus A16 type VP1 protein monoclonal antibody

[0043] (1) Monoclonal antibody ascites preparation:

[0044] Mice: SPF grade mice, no rodent virus contamination after inspection, if the animals are found to be unhealthy, bitten or infected during the ascites production process, they should be discarded.

[0045] Expanded culture of cell lines: take one tube of the production batch of cells to resuscitate, add nutrient solution to expand the culture, one production cell is only used once, and will not be frozen.

[0046]Inoculation of hybridoma cell lines: preparation of ascites should be carried out under sterile conditions, before injection of hybridoma cells, each mouse was intraperitoneally injected with liquid paraffin 0.5ml. One week later, each mouse was intraperitoneally injected with hybridoma cells 1-3×10 6 / 0.2ml.

[0047] Ascites collection: 7-10 days after the injection of the cell line, or the ascites was collected ...

Embodiment 3

[0056] Embodiment 3: Coxsackie virus A16 type (CA16) detection test strip (colloidal gold) (see Fig. 1)

[0057] (1) Preparation of colloidal gold-VP1 monoclonal antibody conjugate:

[0058] It was determined by experiments that the optimum binding pH value of the anti-VP1 monoclonal antibody colloidal label was 8.0, and the ratio of colloidal gold and antibody was 40 μg / ml colloidal gold. After the labeled colloidal gold is treated with a stabilizer, the colloidal gold-antibody conjugate solution is taken in an amount of 65 μl per square centimeter, evenly adsorbed on glass fibers, freeze-dried, and stored in a dry environment.

[0059] (2) Coating antigen on nitrocellulose membrane:

[0060] Dilute another monoclonal antibody of VP1 that recognizes different antigenic sites (please confirm) with 0.01MPBS to 1.5mg / ml. Anti-mouse IgG polyclonal antibody was diluted to 2mg / ml with 0.01MPBS. Spray the two on the nitrocellulose membrane at a speed of 1 μl / cm with a film sprayi...

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Abstract

The invention provides a fast detection testing strip for detecting coxsackie virus A16 (CA16), which is monoclonal antibody or polyclonal antibody and quality control anti-mouse IgG of a nitric acid fiber membrane (NC membrane) coated with coxsackie virus A16 (CA16) specific antigen VP1. The monoclonal antibody combined with coxsackie virus A16 (CA16) specific antigen VP1 labeled by colloidal gold is used for detecting the coxsackie virus A16 (CA16) in the sample by using a membrane chromatography double antibody sandwich method. The application of the detection testing strip has the advantages of simple operation, convenience, rapidity, no special apparatus requirement, no professional training requirement, clear results, easy distinction and easy popularization, thereby the detection testing strip is suitable for basic level, large batch on-site detection for the emergent events and diagnosis at early stage, and has auxiliary effect to the infection diagnosis of coxsackie virus A16 (CA16).

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a Coxsackievirus A16 (CA16) antigen detection test strip and an application thereof. Background technique [0002] Coxsackievirus (Coxsackievirus) is a serious pathogenic agent of human diseases. Its infection can lead to many diseases, from mild respiratory tract infection diseases to more serious myocarditis, pericarditis and some diseases of the nervous system, and even cause infant death. CoxV can be divided into A and B groups: Group A virus has 24 serotypes, and group B has 6 serotypes. All types of group B and type 9 of group A share a common group-specific antigen; there are cross-reactions among various virus serotypes in group B, but not in group A. The CoxV that infects the human body is mostly in group A, and the CoxV that causes hand, foot and mouth disease is mostly A16, A4, A5, A7, A10, B2, B5, and B13. CoxV is a single-stranded positive-strand RN...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569G01N33/532
Inventor 刘明
Owner BEIJING ZHUANGDI HAOHE BIOMEDICINE SCI & TECH
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