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Kit and application thereof

A technology for kits and detection reagents, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., and can solve problems such as cumbersome operation process, high price, and long time consumption

Inactive Publication Date: 2016-08-24
LUOYANG PULIKE WANTAI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the methods for detecting these viruses include virus isolation and identification and serum examination, but the operation process is cumbersome and time-consuming; PCR / RT-PCR detection, but requires specialized instruments and professional operators, these methods are not suitable Grassroots or on-site rapid detection
The prior art also has a single colloidal gold detection test strip, but the price is expensive or the detection sensitivity is low

Method used

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  • Kit and application thereof
  • Kit and application thereof
  • Kit and application thereof

Examples

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Embodiment approach

[0021] As an embodiment of the present invention, the monoclonal antibody 3 is an anti-canine parainfluenza virus monoclonal antibody 4F3B6, and the monoclonal antibody 4 is an anti-canine parainfluenza virus monoclonal antibody 4C9D8.

[0022] As an embodiment of the present invention, the kit further contains effective amounts of anti-canine adenovirus type I monoclonal antibodies 5 and 6, and / or effective amounts of anti-canine adenovirus type II monoclonal antibodies 7 and 8, Wherein, the anti-canine adenovirus type I monoclonal antibodies 5 and 6 are respectively selected from two of C8, E9, G3, C2, H2, and 2E10-H2, and the anti-canine adenovirus type I monoclonal antibody 5 , 6 can react with canine adenovirus type I simultaneously; the anti-canine adenovirus type II monoclonal antibody 7 is anti-CAV-2-Santa from Santa Company, and monoclonal antibody 8 is 4H1-A7.

[0023] Monoclonal antibodies C8 and E9 are disclosed in the preparation and identification of biological c...

Embodiment 1

[0050] Embodiment 1 Preparation, purification, identification and inspection of anti-canine distemper virus monoclonal antibody

[0051] 1.1 Preparation and purification of anti-canine distemper virus monoclonal antibody

[0052] The canine distemper virus CVCC AV299 strain was prepared according to the operating method of Harlow E et al. (Harlow E, Lane D. Antibodies: a laboratory manual. New York: Cold Spring Harbor Laboratory Press, 1998, 139-312). The 6E11 strain and the mouse bone marrow hybridoma cell 1G5 strain were submitted for preservation; the anti-canine distemper virus monoclonal antibody 6E11 and the anti-canine distemper virus monoclonal antibody 1G5 were further prepared by using these two mouse bone marrow hybridoma cell lines . And for the monoclonal antibodies 6E11 and 1G5 respectively according to Chen Dan et al. 8108) was purified by caprylic acid-ammonium sulfate combined precipitation method.

[0053] 1.2 Identification of monoclonal antibodies agains...

Embodiment 2

[0096] Preparation and method, quality research and application of embodiment 2 kit

[0097] 2.1 Colloidal gold detection test strip of the kit

[0098] 2.1.1 Preparation and application of the colloidal gold detection test strip of the kit

[0099] The kit includes colloidal gold detection test strips, sample processing solution (i.e. pH7.4 phosphate buffer containing 2% CHAPS), sample processing tube, sample preservation solution, sample preservation solution (i.e. pH7.4 phosphate buffer solution) in the sample preservation bottle, wherein the preparation of the colloidal gold detection test strip is as follows:

[0100] Prepare colloidal gold solution according to the colloidal gold preparation method that China CN101614737A establishes, and mark respectively monoclonal antibody 2 (the anti-canine distemper virus monoclonal antibody 1G5 that embodiment 1 prepares, dilute to labeling concentration with pH7.4 phosphate buffer saline 350 μg / ml), monoclonal antibody 4 (anti-c...

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Abstract

The invention provides a detection kit and an application thereof and belongs to the field of biological detection. The kit comprises effective doses of anti-CDV (canine distemper virus) monoclonal antibodies 1 and 2, effective doses of anti-CPIV (canine parainfluenza virus) monoclonal antibodies 3 and 4 as well as a detection reagent for detecting an antigen-antibody reaction, wherein the monoclonal antibodies 1 and 2 as well as the monoclonal antibodies 3 and 4 can be subjected to the antigen-antibody reaction with CDV and CPIV respectively at the same time. The kit can effectively detect the two viruses and antigens of the two viruses, when enzyme reaction is adopted for detection, the sensitivity of the enzyme reaction is equivalent to that of PCR (polymerase chain reaction), and when colloidal gold is adopted for detection, the sensitivity of the colloidal gold is better that that of commercially available similar products. The invention further provides the application of the kit in the aspect of non-diagnostic purpose.

Description

technical field [0001] The invention belongs to the field of biological detection, and relates to a kit and an application thereof. Background technique [0002] Canine distemper virus (CDV) belongs to the Morbillivirus genus of the Paramyxoviridae family. It is an RNA virus with a single-stranded negative strand and no segment. Canine distemper (commonly known as canine distemper or measles). Canine distemper was first reported in 1905. It is an acute and highly contagious infectious disease of dogs that occurs widely in the world. It is prone to mixed infection or secondary infection of bacteria and other viruses. The incidence rate is as high as 80%. Afterwards, sequelae such as paralysis, convulsions, and seizures are prone to occur. [0003] Canine parainfluenza virus (CPIV), also known as Parainfluenza virus 5, belongs to the Paramyxovirus genus of the Paramyxoviridae subfamily and is a single-stranded negative-sense RNA virus. The virus is distributed worldwide and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/56983
Inventor 田克恭王莹
Owner LUOYANG PULIKE WANTAI BIOTECH
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