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Muscovy duck parvovirus subunit vaccine

A technology of Muscovy duck parvovirus and subunit vaccine, applied in the direction of viruses, antiviral agents, virus antigen components, etc., can solve the problems of virus spread, parvovirus difference, harm, etc., and achieve the prevention of muscovy duck and gosling plague virus infection , the effect of increasing the antibody level

Inactive Publication Date: 2016-11-23
青岛真源生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disease first appeared in China and many European countries in the middle and late 1960s. It was not called goose parvovirus infection until 1978. The isolated parvoviruses were significantly different
If the dead Muscovy ducks are not handled properly, the virus will often spread and cause greater harm.

Method used

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  • Muscovy duck parvovirus subunit vaccine
  • Muscovy duck parvovirus subunit vaccine
  • Muscovy duck parvovirus subunit vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Embodiment 1, the screening of Muscovy duck parvovirus VP protein

[0015] In 2014, typical symptoms of Muscovy duck parvovirus disease appeared in young Muscovy ducks in a certain duck farm. However, the infected ducks had been vaccinated against Muscovy duck parvovirus before. Therefore, it was suspected that the pathogenic bacteria had a genetic mutation. The immune effect of the vaccine is not good. In order to isolate the source of the disease, the liver, spleen and pancreas of the dying duck were aseptically collected, homogenized with sterile saline to make a suspension, centrifuged to remove the bacteria, and then inoculated with 11-day-old Muscovy duck embryos through the allantoic cavity, and hatched for 168 After 24 hours, the allantoic fluid and embryo body tissue of the dead embryos were collected, homogenized, frozen and thawed repeatedly, and the supernatant was frozen for storage. After the harvested virus liquid was purified, the virus content, immunog...

Embodiment 2

[0018] The construction of the recombinant plasmid that embodiment 2 expresses VP gene

[0019] According to the VP gene sequence and the multiple cloning restriction site of the expression vector PET32a (+), DNAStar software was used to design primers to amplify the full sequence of the VP gene, wherein the 5' end of the forward primer added a BamHI restriction site; A NotI restriction site was added to the 3' end of the reverse primer. The GPV201401 virus was used as a template for PCR amplification, and the target gene was recovered with a DNA purification and recovery kit. Use the pET32a(+) plasmid to obtain the pET32a-VP recombinant plasmid, transfer the recombinant plasmid into expressive Escherichia coli host bacteria, pick a single clone, identify positive clones by enzyme digestion, shake the bacteria, and sequence.

Embodiment 3

[0020] Example 3. Expression, purification, renaturation and SDS-PAGE identification of recombinant fusion protein

[0021] The single cloned recombinant plasmid with correct sequencing results was transformed into expressive Rosetta host bacteria to construct recombinant genetically engineered bacteria. The recombinant genetically engineered bacteria were inoculated into LB bacterial medium containing ampicillin (100mg / L), cultured overnight at 37°C with shaking (200r / min), and the next day, 1ml of the culture was taken out and inoculated into 120ml of LB medium, 37 After culturing at ℃ for 2.5 hours with shaking (200r / min), the bacterial solution was shaken to a translucent and semi-turbid state, and the OD600 was measured by absorbance photometry=0.6~0.8. Before induction, 10ml was taken as a control before induction, and the shaking temperature was changed to Add α-lactose (final concentration: 1.0mmol / L) for induction at 30°C, collect 10ml of bacterial liquid for 1h, 2h, ...

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Abstract

The invention aims to provide a muscovy duck parvovirus subunit vaccine. The muscovy duck parvovirus subunit vaccine comprises an antigen and a vaccine adjuvant, wherein the used antigen is muscovy duck parvovirus VP protein; and the amino acid sequence of the muscovy duck parvovirus VP protein is SEQ ID NO: 1. After muscovy ducks are inoculated with the vaccine prepared by the invention, the antibody level of the muscovy ducks can be improved, the maternal antibody level of filial generation is guaranteed, and young muscovy duck goose parvovirus infection caused by muscovy duck source goose parvovirus is prevented. If young muscovy ducks at the age of one day are inoculated with the vaccine, the young muscovy duck goose parvovirus infection caused by muscovy duck source goose parvovirus can be prevented effectively.

Description

technical field [0001] The invention belongs to the technical field of poultry vaccine preparation, and in particular relates to an inactivated vaccine of muscovy duck parvovirus subunits. Background technique [0002] Parvovirus disease is a highly contagious disease of geese and muscovy ducks (also known as tumor-headed ducks). Raw myocarditis or goose ascites hepatonephritis. According to the age of the infected goslings, the disease can be divided into three types: acute, subacute or chronic. The acute type can cause 100% death of goslings within 10 days of age. Gosling plague virus is mainly transmitted through the secretions, excreta, breeding eggs of sick geese, and water, feed, utensils, and venues that sick geese have drunk. Clinical manifestations include mental fatigue, lone spouse, serous nasal discharge from the nostrils, frequent shaking of the head of the affected goose, loose grayish-yellow or yellow-green feces, nervous disorders, necrosis and shedding of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/23A61P31/20
CPCA61K39/12A61K2039/555A61K2039/55505C12N2750/14034A61K39/23A61P31/20Y02A50/30
Inventor 类延乐张铭李明亮沈彤孙翠彦
Owner 青岛真源生物技术有限公司
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