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Reagent kit for detecting syncytial virus of respiratory passage

A technology for detecting kits and syncytial virus, applied in the direction of measuring devices, biochemical equipment and methods, instruments, etc., to achieve the effect of convenient storage and transportation, and convenient clinical and home use

Inactive Publication Date: 2008-02-27
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

[0026] It can be seen from the existing detection methods of respiratory syncytial virus that although virus isolation, EIA, and RT-PCR diagnostic methods have the advantages of certain specificity and sensitivity, they need professional and technical personnel, special equipment and equipment in operation. Specific conditions and time-consuming shortcomings, 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 a new development direction for syncytial virus diagnosis

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: Preparation of anti-respiratory syncytial virus monoclonal antibody

[0042] (1) Myeloma cells

[0043] 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.

[0044] After the Hep-2 cells are cultured, they are directly spread on glass slides and stored in a -30°C ultra-low temperature refrigerator for monitoring anti-cell clones.

[0045] (2) Immune parental cells

[0046] Respiratory syncytial virus for immunization was purchased from the Institute of Virology, Chinese Academy of Preventive Medicine. The virus strain was inoculated in Hep-2 cells and cultured, and when the cytopathic effect reached (+++), it was immediately stored in a -70°C ultra-low temperature refrigerator, and used as an antigen for immunized anim...

Embodiment 2

[0062] Example 2: Examination of murine virus

[0063] In order to identify the specificity of the prepared anti-RSV monoclonal antibody, that is, no cross-reaction with the used murine virus.

[0064] Check for murine viruses including: hemorrhagic fever virus (EHFV); lymphocyte choroidal meningitis virus (LCMV); type 3 reovirus (Reovirus); Sendai virus; Murine pneumonia virus (PVM).

[0065] Cell inoculation method: Inoculate the secreted supernatant of the 2A7 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.

[0066] Animal inoculation method: 2A7 hybridoma cell line inoculates 10 suckling mice within 24 hours after birth; 10 adult mice each weighin...

Embodiment 3

[0067] Embodiment 3: Mycoplasma inspection

[0068] Culture method: 2A7 hybridoma cell line cell preparations were added with 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. Result: negative control result (-); 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 hybridomas cell strain with preserving number at CGMCC 1546, anti-respiratory syncytial virus N protein monoclone antibody and respiratory syncytial virus detecting agent box (colloidal gold method), which can detects the respiratory syncytial virus with high specificity, sensibility and accuracy to be preserved and transported conveniently for clinical and domestic usage with industrial property.

Description

technical field [0001] The invention relates to an anti-respiratory syncytial virus monoclonal antibody, a hybridoma cell line producing the monoclonal antibody and a respiratory syncytial virus detection kit containing the monoclonal antibody. Background technique [0002] Respiratory syncytial virus (RSV, referred to as syncytial virus, belonging to Paramyxoviridae) was first isolated in 1956 from orangutans with cold symptoms. It is the most common pathogen causing viral pneumonia in children. It is named because it can form special cell fusion lesions in cell culture. Infection with the virus can cause interstitial pneumonia and bronchiolitis. In Beijing, 48% of viral pneumonia and 58% of bronchiolitis were caused by syncytial virus (1980-1984); in Guangzhou, 31.4% of children's pneumonia and bronchiolitis were caused by syncytial virus (1973-1986) ; In the United States, 20% to 25% of infant pneumonia and 50% to 75% of bronchiolitis are caused by syncytial virus. [...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/18G01N33/577G01N33/569
Inventor 王炳彦
Owner 北京阿斯可来生物工程有限公司
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