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A kind of anti-goat pox virus vhh-4-2 based on Bactrian camel and its application

A goat pox virus and gene technology, applied in the field of biology, can solve the problems of high cost, labor, low accuracy and sensitivity

Active Publication Date: 2020-06-09
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problems of time-consuming, laborious, high cost, and low accuracy and sensitivity in the prior art using primary and secondary antibodies, the present invention has developed a key marker against sheeppox virus and provided a dual-based Camelia anti-goat pox virus single domain antibody VHH-4-2

Method used

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  • A kind of anti-goat pox virus vhh-4-2 based on Bactrian camel and its application
  • A kind of anti-goat pox virus vhh-4-2 based on Bactrian camel and its application
  • A kind of anti-goat pox virus vhh-4-2 based on Bactrian camel and its application

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Experimental program
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Effect test

Embodiment

[0064] Screening of a Bactrian camel anti-goat pox virus single domain antibody VHH-4-2 gene

[0065] 1.1 Use camel-derived single-domain antibody homology arm primers to amplify the single-domain antibody gene from the cDNA antibody library (see publication number 201610281583.9), 900bp is the VH-CH1-CH2 hinge region, 600bp is the VHH-CH2 hinge region, and the VHH is about 400bp, 900bp and 600bp bands were recovered from the gel respectively, and 400bp were amplified on this basis. After the gel was recovered, it was connected to the T carrier, transformed into JM109 competent cells, picked spots, shaken bacteria, identified, and verified the appropriate sequence for sequencing . 50ul reaction system: 25ul Ex Taq mix 25ul, targeted upstream and downstream primers 1ul each, template 1ul (cDNA library plasmid), ddH 2 O22ul, amplification conditions: 95°C for 3min, 95°C for 50s, 54°C for 50s, 35times, 72°C for 50s. The conditions of the three fragments are consistent. 1.2 The...

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Abstract

The invention discloses a bactrian camel-based anti-goat pox virus VHH-4-2 and application thereof, and belongs to the field of biotechnology. The technical scheme includes: screening, expressing, purifying, functional labeling and activity verifying of anti-goat pox virus VHH-4-2, concentrating and purifying of goat pox virus, optimizing of various indicators of a kit, etc. The functionally labeled VHH-4-2 is used in the invention, thus avoiding operation steps of primary antibody, secondary antibody, anti-antibody, etc., improving sensitivity and stability, shortening time and saving resources. The bactrian camel-based anti-goat pox virus VHH-4-2 breaks through shortcomings that the traditional Elisa method is time-consuming, laborious, etc. The bactrian camel-based anti-goat pox virus VHH-4-2 lays a foundation for establishing a clinical diagnosis method for sensitive goat pox virus and for studying the infection mechanism of the virus; and the bactrian camel-based anti-goat pox virus VHH-4-2 also provides a key marker for the development of a goat pox virus antibody-immobilized competitive Elisa kit with complete proprietary intellectual property rights. In addition, the attenuated goat pox vaccine is used as a coating antigen after expanded culture, concentration and purification, so that the sensitivity is improved.

Description

technical field [0001] The invention belongs to the field of biology and relates to a Bactrian camel-derived anti-goat pox virus VHH-4-2, in particular to a Bactrian camel-derived anti-goat pox virus VHH-4-2 and its use. Background technique [0002] Sheep pox is an acute, febrile, highly contagious infectious disease caused by sheep pox / goat pox virus or mixed infection, causing fever, papules or herpes on the skin and mucous membranes of sheep and goats with less or no hair. It is the most serious of all animal pox diseases. Because of the similarity of clinical symptoms with sheep contagious pustule (commonly known as sheep mouth sore), the crossover of serological reactions, and the frequent occurrence of visceral type sheep pox have caused greater difficulties in the diagnosis of the disease. [0003] Single-domain antibody (VHH) is currently available as the smallest structural unit with stable structure, complete function, and ability to bind antigens. Due to its low...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/08C07K1/34G01N33/535G01N33/569
CPCC07K16/081C07K2317/22C07K2317/569G01N33/535G01N33/56983G01N2333/065
Inventor 吴锦艳尚佑军田宏尹双辉曹小安王耀杰惠小婷关玉华刘湘涛刘永生
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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