Porcine coronavirus vaccines

A coronavirus, virus technology, applied in the field of combined vaccines that provide both PEDV and PDCoV antigens, can solve problems such as protection or reduction of clinical symptoms of unproven diseases

Pending Publication Date: 2019-12-06
BOEHRINGER INGELHEIM ANIMAL HEALTH USA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, although Marx et al. also described a D-coronavirus vaccine, no protection or reduction in clinical signs of disease was demonstrated with monovalent PDCoV vaccines or bivalent PEDV and PDCoV vaccines.

Method used

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  • Porcine coronavirus vaccines
  • Porcine coronavirus vaccines
  • Porcine coronavirus vaccines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0183] Embodiment 1: Isolation and production of PEDV strain

[0184] To produce a porcine epidemic diarrhea virus vaccine, the virus is killed and a stock culture of PEDV (isolate) is first produced. PEDV cultures were grown from this stock seed and then inactivated. The inactivated virus culture is then mixed with an adjuvant in order to produce a porcine epidemic diarrhea virus vaccine. The following method was used for the production of porcine epidemic diarrhea virus vaccine.

[0185] Animals or tissues from animals exhibiting extreme diarrhea were obtained in 2013. Homogenates from mucosal scrapings were generated from these animals, filtered through a 0.2 micron syringe filter, and the filtrate was used to inoculate Vero cells (VERO). Viruses were grown in the presence of PEDV maintenance medium containing modified MEM, porcine trypsin, tryptone phosphate broth, yeast extract and HEPES buffer. Viral growth was assessed and visualized by examining characteristic sync...

Embodiment 2

[0191] Embodiment 2: Isolation and production of porcine D-coronavirus strain

[0192] Porcine D-coronavirus (PDCoV) was purchased from National Veterinary Service laboratory (NVSL), Ames, IA. The virus was isolated at NVSL from pig intestine samples from Illinois in 2014. Virus was isolated on porcine testicular (ST) cells in serum-free medium in the presence of 5 μg / mL TPCK trypsin. The original virus was subjected to two rounds of plaque purification at NVSL, and the tenth passage of virus was used as starting material.

[0193] Passage 10 virus was inoculated and adapted to porcine testicular (ST) cells at BI, maintained with PDCoV containing modified minimal essential medium with HEPES, tryptone phosphate broth, yeast extract, and porcine trypsin base. Cells were incubated at 37°C with 5% CO2 for 24-48 hours and observed for evidence of PDCoV-induced cytopathic effects. When CPE was complete and cells had detached, flasks were freeze-thawed once and lysates were harve...

Embodiment 3

[0194] Example 3: Genome sequence analysis of PEDV isolates and PDCoV isolates

[0195] Sample preparation and analysis:

[0196] Prior to extraction, viral tissue culture supernatants are pretreated with a mixture of DNase and RNase to remove residual host cell genomic nucleic acid. then use Viral RNA Extraction Kit (Qiagen, catalog #52906), for the extraction of viral genomic RNA from nuclease-treated samples. After extraction, the samples were again treated with DNase to further enrich for viral genomic RNA. Subsequently, viral genomic RNA was converted to double-stranded cDNA (ds cDNA) by randomly primed reverse transcription and Klenow fragment treatment. then use XT Library Prep Kit (Catalog #FC-131-1024), ds cDNA products are used to generate Illumina-based Sequencer for sequencing the library. Each sample is barcoded with unique tags on both the 5' and 3' ends to minimize the chance of mis-binning biological information. The library was published at The se...

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Abstract

The present invention relates to a vaccine for protecting a pig against diseases associated with corona virus infection including porcine epidemic diarrhea virus (PEDV) and / or porcine deltacorona virus (PDCoV). The vaccine commonly includes inactivated / killed PEDV (e.g., chemically inactivated PED virus), and / or recombinant PEDV antigen, and / or an adjuvant inactivated / killed PDCoV (e.g., chemically inactivated PDCoV virus), and / or recombinant PDCoV antigen and an adjuvant. Methods for protecting pigs against diseases associated with PEDV and / or PDCoV and methods of producing the porcine epidemic diarrhea virus and / or porcine deltacorona virus vaccine are also provided.

Description

[0001] sequence listing [0002] This application contains a Sequence Listing pursuant to 37 C.F.R. 1.821-1.825. The contents of the 480kb ASCII text file of the Sequence Listing titled 10-0176-WO-SEQ were created on January 23, 2017 and are accompanied by electronic filing via EFS-Web, which application is hereby incorporated by reference in its entirety. technical field [0003] The present invention relates to immunogenic compositions that protect pigs from disease caused by specific porcine epidemic diarrhea virus (PEDV) vaccines and porcine D-coronavirus (PDCoV), as well as providing a combination of both PEDV and PDCoV antigens vaccine. Epidemic enteric disease, often co-occurring, is a major economic concern for the US swine industry due to high mortality (up to 100%) in piglets < 10 days of age. Background technique [0004] Porcine epidemic diarrhea virus is an enveloped, positive-sense, single-stranded RNA virus that causes acute diarrhea, vomiting and dehydr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61K39/00
CPCA61K39/12A61K2039/5252A61K2039/552A61K2039/70C12N2770/20034A61P1/00A61P1/12A61P31/14A61P37/04A61P43/00A61K39/215A61K39/39C12N7/00A61K2039/55566C12N2770/20063C12N2770/20051C12N2710/14043C12N2710/14071C12N2770/20071
Inventor S·阿克曼J·R·赫尔曼L·A·赫尔南德斯L·A·霍布斯A·V·耶尔S·奥康纳A·R·帕特森E·M·沃恩J·G·维多利亚
Owner BOEHRINGER INGELHEIM ANIMAL HEALTH USA INC
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