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Influenza Virus A colloidal gold quick detection test paper

A technology for detecting influenza A virus and test strips, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., to achieve the effect of convenient storage and transportation

Inactive Publication Date: 2009-06-24
北京阿斯可来生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0038] It can be seen from the existing influenza detection methods that although virus isolation, EIA, and RT-PCR diagnostic methods have certain advantages in specificity and sensitivity, they require professional and technical personnel, special equipment, and specific conditions and conditions in operation. Time-consuming and other disadvantages, and the mature immune colloidal gold diagnostic technology developed in recent years has high specificity, high sensitivity, simple operation and does not require professionals and equipment, and has become the development direction of rapid influenza diagnosis

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1: Preparation of anti-influenza A virus monoclonal antibody

[0054] (1) Myeloma cells

[0055] SP2 / 0 myeloma cells: purchased from the Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences. Resuscitate the SP2 / O 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.

[0056] After the DMCK cells are cultured, directly smear them on glass slides and store them in a -30°C ultra-low temperature refrigerator for monitoring anti-cell clones.

[0057] (2) Immune parental cells

[0058] Influenza A1 (A / Jingfang 25 / 96H1N1) and Influenza A3 (A / Shanghai Fang / 1 / 98H3N2) viruses were purchased from the National Influenza Center, Institute of Virology, Chinese Academy of Preventive Medicine. The recovered virus strains were respectively inoculated in DMCK cells for culture, and when the cytopathic effect reached (+++), they were immediately stored in ...

Embodiment 2

[0076] Example 2: Examination of murine virus

[0077] In order to identify the specificity of the anti-influenza A virus nucleoprotein monoclonal antibody prepared, that is, there is no cross-reaction with the mouse virus used.

[0078] Check for murine viruses including: hemorrhagic fever virus (EHFV); lymphocytic choriomeningitis virus (LCMV); type 3 reovirus (Reovirus); Sendai virus; Mouse pneumonia virus (PVM).

[0079] Cell inoculation method: Inoculate the secreted supernatant of CGMCC 0987 3F1 hybridoma cell line into Vero (African green monkey kidney passage cells); 2BS (human embryonic lung) diploid cells, the inoculation amount is 10 7 . After the cells were inoculated, they were passed down for two generations, and the cells were observed for lesions. At the same time, slices were made, and the virus antigen was detected by the IFA method, which was negative.

[0080] Animal inoculation method: CGMCC 0987 3F1 hybridoma cell line inoculated 10 suckling mice withi...

Embodiment 3

[0082] Embodiment 3: mycoplasma inspection:

[0083] Cultivation method: CGMCC 0987 3F1 hybridoma cell line cell preparation plus mycoplasma monoclonal antibody (purchased from the Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences) for 30 minutes, fluorescent staining, and negative and positive controls at the same time, the result: negative control results ( -); positive control result (+); test specimen result: negative. After the culture supernatant was coated, monoclonal antibody was added, detected by ELISA method, and a negative control was made at the same time, and the result of the specimen was negative.

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Abstract

The invention discloses a hybridoma cell strain with preservation number CGMCC0987, an anti-influenza A virus nucleoprotein monoclonal antibody produced by the hybridoma cell strain, and a colloidal gold rapid detection test paper for influenza A virus containing the monoclonal antibody. The detection test paper can detect rapid influenza A virus, has high specificity, sensitivity and accuracy, is convenient for storage and transportation, is extremely convenient for clinical and household use, and is industrial.

Description

technical field [0001] The invention relates to an anti-influenza virus nucleoprotein monoclonal antibody, a hybridoma cell line producing the monoclonal antibody and a colloidal gold rapid detection test paper for the influenza A virus containing the monoclonal antibody. Background technique [0002] Influenza (influenza) is called flu for short, is the acute respiratory infectious disease that is caused by influenza virus. The clinical features are rapid onset of high fever, body aches, fatigue, or mild respiratory symptoms. The disease has a short incubation period, is highly contagious, and spreads rapidly. Influenza viruses are divided into three types: A, B, and C, and type A is the most threatening. Due to the strong pathogenicity of influenza virus, it is easy to mutate, and the population lacks immunity to mutant strains, which can easily cause outbreaks. Influenza is very harmful to humans, and an influenza epidemic can lead to a reduction in the average life exp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/06G01N33/53G01N33/543G01N33/569G01N33/577
Inventor 王炳彦
Owner 北京阿斯可来生物工程有限公司
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