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ELISA detection kit for canine distemper and canine parvovirus antibody

A detection kit and technology for canine distemper, which is applied in the field of ELISA detection kits, can solve the problems of cumbersome work, increased workload, and unfavorable human health, and achieve the effect of convenient use and reduced use cost

Inactive Publication Date: 2017-08-04
YANGLING LVFANG BIOLOGICAL ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] my country's dog disease prevention and control lacks an effective comprehensive prevention and control system, which is not conducive to the prevention and control of dog diseases, nor is it conducive to human health
There is often a situation in dogs after vaccination, that is, the immune effect is not known, which leads to some dogs still getting sick after vaccination, and the popular kits on the market can only detect canine distemper or canine parvovirus. Common diseases usually require the purchase of 2 kits for detection, which virtually increases the workload and is cumbersome

Method used

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  • ELISA detection kit for canine distemper and canine parvovirus antibody

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Experimental program
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Effect test

preparation example Construction

[0034] 1. Preparation of purified canine IgG:

[0035] The dogs that passed the inspection were selected as dogs for blood collection. Before blood collection, they were in good condition, with normal body temperature, appetite, urine and stool, and no other diseases. After disinfection, blood was collected from the jugular vein, serum was separated, and the IgG purification scheme was caprylic acid-ammonium sulfate precipitation method.

[0036] Take 1 part of pretreated serum and add 2 parts of 0.06mol / L acetate buffer solution with pH 5.0, adjust the pH to 4.8 with 1mol / L HCl; add 11ul caprylic acid per ml of diluted serum, add dropwise under stirring at room temperature Caprylic acid, add it within 30 minutes, let it stand at 4°C for 2 hours, take it out and centrifuge at 12000r / min for 30min, discard the precipitate; filter the supernatant through a nylon sieve with a pore size of 125um, add 1 / 10 volume of 0.01mol / L PBS, adjust the pH to 7.2 with 1mol / L NaOH solution; a...

example 2

[0081] 1. Antigen preparation:

[0082] 1. Expression of canine distemper virus H protein and canine parvovirus VP2 gene:

[0083] According to the H gene sequence of distemper virus and the VP2 gene sequence of canine parvovirus in GenBank, two pairs of primers were designed and synthesized, the H gene fragment and the VP2 gene fragment were amplified from the DNA by PCR method, and cloned into the expression vector pET28b to construct The recombinant plasmids pET-H and pET-VP2 genes were obtained, and after sequencing and verification, they were transformed into the expression host strain BL21(DE3), and their expression was induced by IPTG.

[0084] 2. Purification of canine distemper virus H protein and canine parvovirus VP2 gene:

[0085] After the induction, collect the induced cells by centrifugation, wash the resuspended cells with PBS, and ultrasonically lyse. The specific process is: ultrasonic for 1.0 s, with an interval of 1 s, for a total of 10 min; then centrifug...

example 3

[0134] 1. Antigen preparation:

[0135] 1. Expression of canine distemper virus H protein and canine parvovirus VP2 gene:

[0136] According to the H gene sequence of distemper virus and the VP2 gene sequence of canine parvovirus in GenBank, two pairs of primers were designed and synthesized, the H gene fragment and the VP2 gene fragment were amplified from the DNA by PCR method, and cloned into the expression vector pET28b to construct The recombinant plasmids pET-H and pET-VP2 genes were obtained, and after sequencing and verification, they were transformed into the expression host strain BL21(DE3), and their expression was induced by IPTG.

[0137] 2. Purification of canine distemper virus H protein and canine parvovirus VP2 gene:

[0138] After the induction, collect the induced cells by centrifugation, wash the resuspended cells with PBS, and ultrasonically lyse. The specific process is as follows: ultrasonic 1.0s, interval 2s, a total of 15min; then centrifuge at 12000r...

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Abstract

The invention discloses an ELISA detection kit for a canine distemper and canine parvovirus antibody. The ELISA detection kit comprises an ELISA slat, a serum diluent, a concentrated detergent, a substrate developing solution, a stop solution, an enzyme-labelled monoclonal antibody, a positive control, a negative control, and a horseradish peroxidase conjugated-rabbit anti-dog IgG polyclonal antibody. The ELISA detection kit for the canine distemper and canine parvovirus antibody is convenient and efficient, and a serum antibody is detected through serum to judge the canine immune protection condition after vaccination.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to an ELISA detection kit for canine distemper and canine parvoantibodies. Background technique [0002] In our country, with the transformation of social and economic development mode, the scale of dog breeding, fur animal breeding and wild animal protection industry is constantly expanding; with the continuous improvement of people's quality of life, the number of pet dogs is also increasing year by year , and the biggest impact on the development of these industries is the outbreak of canine infectious diseases, and the prevention and control of these infectious diseases are becoming more and more severe. Among canine diseases, canine distemper and canine parvo are diseases with high morbidity and mortality. Canine distemper is a kind of highly contagious disease that is caused by canine distemper virus, and infectiousness is extremely strong, and the death rate can reach more than ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56983G01N2333/13
Inventor 袁鹏飞焦铁军李河林胡江锋
Owner YANGLING LVFANG BIOLOGICAL ENG CO LTD
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