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Triple inactivated vaccine and preparation method thereof

An inactivated vaccine and inactivated technology, which is applied in the direction of botany equipment and methods, biochemical equipment and methods, vaccines, etc., can solve the problems of virus protection that cannot be mutated, and achieve the improvement of chicken flock protection rate, long duration, Good safety performance

Inactive Publication Date: 2020-08-07
扬州优邦生物药品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There are currently no vaccines against the novel IBDV variants and AI variants, and traditional vaccines do not provide protection against circulating variants of the virus

Method used

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  • Triple inactivated vaccine and preparation method thereof
  • Triple inactivated vaccine and preparation method thereof
  • Triple inactivated vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1 3

[0063] The antigen composition of embodiment 1 triple inactivated vaccine

[0064] The antigen of the triple inactivated vaccine is the VP2 protein of the inactivated Newcastle disease La Sota virus strain, the inactivated H9 subtype avian influenza TL18 strain and the inactivated variant strain bursal virus FJ19; the variant strain bursal virus VP2 The nucleotide sequence of the protein is shown in SEQ ID NO.1; the amino acid of the mutant strain bursal virus VP2 protein is shown in SEQ ID NO.2; the content of the chicken Newcastle disease La Sota virus strain ≥ 10 8.5 EID 50 / 0.1mL; the content of the H9 subtype avian influenza TL18 strain ≥10 7.5 EID 50 / 0.1mL; the agar expansion titer of the VP2 protein content of the variant strain bursal virus ≥ 1:128;

[0065] The chicken Newcastle disease La Sota virus strain is derived from China Veterinary Medicine Supervision Institute;

[0066] The H9 subtype avian influenza TL18 strain belongs to influenza virus. The TL18 stra...

Embodiment 2

[0068] The preparation of embodiment 2 bursal virus mutant strain FJ19 strain total cDNA

[0069] Collect the bursa tissue of the diseased chicken, grind it, add double-antibody-containing PBS to dilute it into a suspension, add an equal volume of chloroform to the suspension, place it in a low-temperature environment at 20 rpm for 24 hours, and take the supernatant after centrifugation at 3000 rpm Packed in ampoule vials for later use. The processed samples were inoculated into 10-day-old chicken embryos with the choriourin pathway (CAM), and incubated at 37°C. The chicken embryos that died within 24 hours were discarded, and the chicken embryos that died after 24 hours were placed at 4°C, the allantoic fluid was collected for hemagglutination test, and the embryo bodies and urea membranes were collected at the same time, which were aseptically ground and then packed in ampoules for later use. The total RNA of the culture was extracted using the AXYGEN kit, and the RNA was r...

Embodiment 3

[0070] The preparation of embodiment 3 variant strain bursal virus VP2 protein:

[0071] Preparation of S201, pMD-IBDV-VP2 recombinant plasmid:

[0072]Design primer pairs suitable for expressing the VP2 protein nucleic acid sequence in the baculovirus expression system, named respectively as VP2-P1 and VP2-P2, the nucleotide sequence of the VP2-P1 is shown in SEQ ID NO.3; the VP2 The nucleotide sequence of -P2 is shown in SEQID NO.4; the reverse transcription product cDNA of the total RNA of the extracted mutant strain IBDV FJ19 strain is used as a template, and VP2-P1 and VP2-P2 are used as primers for PCR amplification. The target fragment of VP2 was obtained, and after detection by agarose gel electrophoresis, the target fragment of VP2 was recovered, and the result was as follows figure 2 As shown, among them, M: DL2000 DNA Marker; 1: PCR product, it can be seen from the figure that the size of the recovered fragment is 1356bp, and it is connected to the pMD vector with...

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Abstract

The invention provides a variant bursal disease virus FJ19 strain, a method for preparing a triple inactivated vaccine by using VP2 protein of the strain, and the corresponding vaccine. The vaccine comprises the VP2 protein derived from the variant FJ19 strain (with the preservation number of CGMCC NO: 19381), a newcastle disease La Sota virus strain and an H9 subtype avian influenza TL18 strain (with the preservation number of CGMCC NO: 19382). The triple vaccine disclosed by the invention can prevent and treat three diseases by one injection, and is good in immunogenicity, long in duration and good in safety performance; and particularly, the vaccine is high in VP2 protein content and low in formaldehyde and endotoxin content, has a good protection effect on the currently popular FJ19 variant bursal disease virus, is small in immunizing dose and good in immunizing effect, and can effectively improve the protection rate of chicken flocks.

Description

technical field [0001] The invention belongs to the technical field of biological veterinary drugs, and in particular relates to a novel triple inactivated vaccine of Newcastle disease, infectious bursa and avian influenza and a preparation method thereof. Background technique [0002] Newcastle disease (ND) is an acute, febrile, septic and highly contagious infectious disease of poultry caused by Newcastle disease virus. It is characterized by high fever, dyspnea, diarrhea, nervous disturbance, and mucosal and serosal hemorrhage. It has a high morbidity and fatality rate and is one of the most serious severe infectious diseases that endanger the global poultry industry. OIE lists it as a class A disease, and it is one of the five first-class animal diseases stipulated in my country's "National Medium and Long-Term Animal Disease Prevention and Control Plan" for priority prevention and control. It is listed as a notifiable animal disease by the World Organization for Anima...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/17A61K39/12A61P31/14A61K39/145A61P31/16C12N15/866C07K14/115C07K14/11
CPCA61K39/12A61K2039/5252A61K2039/70A61P31/14A61P31/16C07K14/005C12N15/86C12N2710/14043C12N2720/10022C12N2760/10034C12N2760/16122C12N2760/16134C12N2760/18122C12N2760/18134
Inventor 杨振钱钟王建国李琛马玉峰董昌海杨玉倩江媛徐萍李甜甜范娟潘杰
Owner 扬州优邦生物药品有限公司
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