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Porcine transmissible gastroenteritis virus and application thereof

A technology for transmissible gastroenteritis, applied in porcine transmissible gastroenteritis virus and its application fields, can solve problems such as economic loss, unsatisfactory effect, slow growth, etc., and achieve good safety and good immune protection effect Effect

Active Publication Date: 2014-04-23
兆丰华生物科技(南京)有限公司 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Pigs over 5 weeks old have a low mortality rate, but due to the impact of diarrhea, slow growth and low feed remuneration, resulting in serious economic losses
There are reports of the use of vaccines at home and abroad, the vaccination routes are different, and the effect is not very satisfactory

Method used

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  • Porcine transmissible gastroenteritis virus and application thereof
  • Porcine transmissible gastroenteritis virus and application thereof
  • Porcine transmissible gastroenteritis virus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Isolation and identification of porcine transmissible gastroenteritis virus

[0021] 1. Isolation of porcine transmissible gastroenteritis virus

[0022] A pig farm in Hebei sent the small intestines of pigs with diarrhea for inspection. The contents were collected and tested by RT-PCR. The result was positive for porcine transmissible gastroenteritis.

[0023] Take an appropriate amount of the small intestine submitted for inspection, scrape the intestinal mucosa and contents, add PBS according to the ratio of 1:5 (weight: volume), freeze and thaw repeatedly 3 times, centrifuge to get the supernatant, filter with a 0.22 μm filter membrane, and add the final concentration to the filtrate 20 μg / ml trypsin, 37°C for 1.5 hours.

[0024] Inoculate ST cells with a single layer according to the conventional method (wash three times with PBS of pH 7.4 before inoculation), inoculate the virus at a ratio of 10%, absorb at 37°C for 1 hour, and supplement the cell maint...

Embodiment 2

[0037] Example 2 Attenuation of porcine transmissible gastroenteritis virus

[0038] 1. Attenuated cultivation of porcine transmissible gastroenteritis virus

[0039] The isolated porcine transmissible gastroenteritis virus was continuously passaged on ST cells for 120 generations, and cloned and purified during this process, and then continued to be passed on ST cells for 165 generations. The 165th generation virus was detected by RT-PCR, and the amplified S gene fragment was 886bp.

[0040] The TGEV ORF3 gene sequence was relatively stable in the 20th, 40th, 70th, and 100th generations, with individual base changes but no amino acid changes; and the ORF3 gene appeared together with the previous spacer sequence at the 125th generation Deletion of 81 nucleotides (30 nucleotides before the start codon and 81 nucleotides after the last 51 nucleotides), stable deletion of 81 nucleotides in the 125th, 145th, and 165th generations (The sequence is shown in SEQ ID No.5). The resu...

Embodiment 3

[0066] Example 3 Safety Evaluation of Porcine Transmissible Gastroenteritis Virus HB08 Strain

[0067] Dilute passage 145 virus to 10 6.0 TCID 50 / ml (10 heads), 10 3-day-old piglets were selected, injected intramuscularly, 1ml each, and observed for 14 days, and 5 piglets in the control group were injected with the same dose of normal saline; in addition, pregnant sows one and a half months before delivery were selected. 10 pigs, each with 20 doses intramuscularly, and 5 sows in the control group were injected with the same dose of saline; the indicators of feed intake, body temperature, diarrhea and abortion of the sows were observed, and the litter size of the sows was counted.

[0068] The results proved that large-dose injection of the virus had no effect on piglets and sows, and no fever, diarrhea, and local inflammation occurred. The sows had reproductive disorders such as abortion and stillbirth. There was no significant difference between the test group and the contr...

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Abstract

The invention discloses a low virulent strain HB08 of a porcine transmissible gastroenteritis virus (TGEV) and an application thereof. The microbial collection number of the low virulent strain is CGMCC No.7807. The low virulent strain of the TGEV is good in safety, and the virulent strain is safe to the pigs of all ages including pregnant sows and piglets. The low virulent strain has 100% active protection rate and 97.2% passive protection rate on three-day-old piglets, therefore, the low virulent strain HB08 of the porcine transmissible gastroenteritis virus is indicated to have excellent immunoprotection effect on the porcine transmissible gastroenteritis.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a strain of porcine transmissible gastroenteritis virus and its application. Background technique [0002] Porcine transmissible gastroenteritis (TGE) is an important contact viral intestinal infectious disease that causes pig diarrhea. The mortality rate of pigs over 5 weeks old is low, but due to the impact of diarrhea, the growth is slow and the feed remuneration is low, causing serious economic losses. The pathogen was identified by Doyle et al. in 1946. Since the late 1960s, the disease has been prevalent in many provinces and cities in my country. [0003] Since the disease became popular, experts at home and abroad have done a lot of work. At present, there is no specific preparation to resist porcine transmissible gastroenteritis virus. Clinically, symptomatic treatment is often used to relieve symptoms and reduce mortality. There are reports of the use of vacc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/225A61P31/14C12R1/93
Inventor 侯艳红王贵华赵亚荣于萍萍刘明明卢会英满坤陈翠云
Owner 兆丰华生物科技(南京)有限公司
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