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Immunochromatographic test strip for detecting feline panleukopenia syndrome virus antibody, preparation method and application thereof

A leukopenia, syndrome technology, applied in the directions of virus/phage, botanical equipment and method, biochemical equipment and method, etc., can solve the monitoring of limited application, high test conditions and operation level, reducing the immune effect of syndrome virus And other issues

Active Publication Date: 2021-04-09
北京纳百生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The hemagglutination inhibition test requires fresh and highly sensitive red blood cells, and the detection time takes 2 to 3 hours; the neutralization test is cumbersome to operate, requires high test conditions and operating levels, and the entire test period is long; the enzyme-linked immunosorbent assay method has high requirements for operation. The results of the microplate reader are required, and it is not suitable for the detection of a single sample
All the inconveniences mentioned above limit their application in the clinical detection of feline parvovirus antibody
[0006] The purpose of the patent of the present invention is to overcome the shortcomings of the above several methods, to establish a simple, sensitive, specific and fast qualitative detection method for feline panleukopenia syndrome virus antibody, and to apply it to the detection of the immune effect of feline panleukopenia syndrome virus Surveillance to limit the prevalence of feline panleukopenia syndrome virus

Method used

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  • Immunochromatographic test strip for detecting feline panleukopenia syndrome virus antibody, preparation method and application thereof
  • Immunochromatographic test strip for detecting feline panleukopenia syndrome virus antibody, preparation method and application thereof
  • Immunochromatographic test strip for detecting feline panleukopenia syndrome virus antibody, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1 Expression, purification and identification of feline panleukopenia syndrome virus VP2 recombinant protein

[0076] 1. Search for the VP2 gene in Genbank, the accession number is: MN451692, and the sequence is shown in SEQ ID NO.1.

[0077] 2. Segmented expression of VP2 recombinant protein

[0078] The truncated peptides of the VP2 protein were expressed in multiple fragments, and the carrier used was PET-32a. Using the method of segmented expression of peptides, the VP2 protein with the signal peptide removed is expressed in 6 segments, so that each peptide segment overlaps (the schematic diagram of the segments is shown in figure 1 shown), the amplification primers for each fragment are as follows:

[0079] Primers for amplifying the P1 fragment:

[0080] F1 5'-caaggatccGGATCTGGGAACGGGTCTGGAG-3' (SEQ ID NO. 2),

[0081] R1 5'-caagtcgacATCTCCTGGATTAAACCAAAC-3' (SEQ ID NO. 3);

[0082] Primers for amplifying the P2 fragment:

[0083] F2 5'-aaaggatccATTG...

Embodiment 2

[0115] Example 2 Preparation of a conjugate of colloidal gold-labeled streptavidin and goat anti-cat anti-antibody-biotin conjugate

[0116] (1) Preparation of streptavidin-colloidal gold markers

[0117] Take 1.5% colloidal gold placed at room temperature, take 1mL with a pipette, put it into a PE tube, add 3μL 0.2M potassium carbonate, shake and mix, and set aside; prepare streptavidin with 0.01M PBS (pH 8.0) to 5mg / ml Take 4 μL of the solution with a pipette and add it to the colloidal gold solution, shake and mix, and let it stand still for 10 minutes; draw 10 μL 10% BSA with a pipette and add it to the colloidal gold solution, shake it and mix it, and let it stand still for 8 minutes; Pipette 10 μL of 10% polyethylene glycol 20000 into the colloidal gold solution, shake and mix; centrifuge at 12,000 r / min for 5 minutes; discard the supernatant, take 100 μL of reconstitution solution to redissolve, and mix well.

[0118] (2) Preparation of goat anti-cat anti-antibody-biot...

Embodiment 3

[0123] Example 3 Preparation of Conjugate Release Pad

[0124] The prepared conjugate of colloidal gold-labeled streptavidin and goat anti-cat anti-antibody-biotin conjugate is evenly sprayed on the conjugate release pad (conjugate release pad) with a Bio dot film-drawing instrument. Spray 1 μL of the conjugate of colloidal gold-labeled streptavidin and goat anti-cat anti-antibody-biotin conjugate on a 1cm conjugate release pad, then place it at 40°C for 16 hours, take it out, and store it in a dry environment spare.

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Abstract

The invention discloses an immunochromatographic test strip for detecting feline panleukopenia syndrome virus antibody, a preparation method and an application thereof. The immunochromatographic test strip prepared by the invention has good sensitivity, specificity and stability, convenient storage, long storage period, simple operation and fast detection speed, and is suitable for rapid diagnosis of feline panleukopenia syndrome virus infection .

Description

technical field [0001] The invention relates to the field of biotechnology, and relates to an immunochromatographic test strip for detecting feline panleukopenia syndrome virus antibody, a preparation method and an application thereof. Background technique [0002] Feline panleukopenia virus (FPV), also known as feline parvovirus, feline infectious enteritis virus or feline distemper virus, can cause acute, highly contagious infectious diseases characterized by high fever, vomiting, enteritis and severe leukopenia as a feature. The virus mainly infects a variety of animals such as cats and mustelidae, and is currently the virus with the widest infection range and the strongest pathogenicity among the carnivorous parvoviruses. And young animals are highly infectious, posing a great threat to wild animals and pet cats. The disease exists in the world, and it occurs in many places in my country, which is extremely harmful to cats and other cats. There are a variety of commer...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/015C12N15/35G01N33/68G01N33/569G01N33/558
CPCC07K14/005C12N2750/14322G01N33/558G01N33/56983G01N33/6854G01N2333/015G01N2469/20
Inventor 于在江吴迪郭秀锋李铁刚张德红袁志波
Owner 北京纳百生物科技有限公司
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