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A kind of attenuated strain ch-p60 of type 3 duck hepatitis A virus and its application

A CH-P60 and duck hepatitis A virus technology, which is applied to the attenuated duck hepatitis A virus type 3 strain CH-P60 and its application fields, can solve problems such as economic losses in the breeding industry, and achieve the effect of wide application value

Active Publication Date: 2021-06-08
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the prevention and control of DHAV in the market mainly uses the attenuated vaccine of DHAV-1. They can effectively prevent and control the prevalence of DHAV-1 in my country, but they cannot completely prevent the prevalence of DHAV-3 viruses belonging to different serotypes from causing damage to the aquaculture industry. huge economic loss

Method used

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  • A kind of attenuated strain ch-p60 of type 3 duck hepatitis A virus and its application
  • A kind of attenuated strain ch-p60 of type 3 duck hepatitis A virus and its application
  • A kind of attenuated strain ch-p60 of type 3 duck hepatitis A virus and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Isolation and Identification of Type 3 Duck Hepatitis A Virus

[0022] 1. Virus Isolation

[0023] The DHAV-3CH isolate was isolated in 2013 from 9-day-old onset Peking ducks in duck farms around Chengdu City, Sichuan Province. The specific process is as follows:

[0024] Aseptically collect the liver of dead ducklings, add sterilized phosphate buffer solution at a ratio of 1:5 by volume, grind, freeze and thaw 3 times, centrifuge at 12000g for 10min, filter the supernatant through a 0.22μm filter and pass through urine Ten 9-day-old duck embryos were inoculated via the cystic cavity route, and each embryo was inoculated with 0.2 mL. The inoculated duck embryos were incubated in a constant temperature incubator at 37°C, and the embryos were photographed once every 8 hours. The death of duck embryos within 7 days of inoculation was recorded. The dead duck embryos within 24 hours were discarded, and the allantoic fluid of the remaining dead duck embryos was col...

Embodiment 2

[0042] Embodiment 2 The cultivation of type 3 duck hepatitis A virus attenuated strain (CH-P60)

[0043] 1. The strain was weakened by continuous passage

[0044] Use 9-day-old duck embryos to continuously passage and attenuate the type 3 duck hepatitis A virus isolate (CH-P10). In each passage, 5 duck embryos are inoculated through the allantoic cavity, 0.2mL / piece, and placed at a constant temperature of 37°C after inoculation. Continue to incubate in the incubator, discard the chicken embryos that died within 24 hours, harvest the duck embryo allantoic fluid of duck embryos that died within 24 to 48 hours, mix well, and then use sterilized physiological saline to dilute the embryo fluid 100 times before proceeding to the next step. One passage, such continuous passage for 50 generations to obtain CH-P60, and finally carry out virus content, sterility and mycoplasma detection, exogenous virus detection and safety test on susceptible ducklings to determine whether the virulen...

Embodiment 3

[0057] Example 3 Application of Type 3 Duck Hepatitis A Virus Attenuated Strain (CH-P60)

[0058] 1. Preparation of virus fluid

[0059] The attenuated strain of duck hepatitis A virus type 3 (CH-P60) obtained in Example 2 was diluted 100 times with sterilized physiological saline as a seed poison, and 20 9-day-old duck embryos were inoculated into the allantoic cavity, 0.2 mL per embryo, and placed at 37 ° C. Continue to incubate in the constant temperature incubator, irradiate the eggs once 24 hours after inoculation, discard the dead embryos, and then irradiate the eggs once every 8 hours, take out the dead duck embryos at any time, until 48 hours, all duck embryos are dead, and put The air cells of the collected duck embryos were erected upright, and cooled at 4°C for 8 hours, and the aseptically collected duck embryo allantoic fluid after cooling was stored at -20°C for future use.

[0060] 2. Examination of venom

[0061] The virus liquid of harvest carries out sterili...

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Abstract

The invention discloses a type 3 duck hepatitis A virus attenuated strain CH-P60 and its application. The virus strain was preserved in the China Typical Culture Collection Center on December 2, 2018. The preservation address is: No. 8, Wuchang District, Wuhan City, Hubei Province On the campus of Wuhan University, No. 299, 1st Road, the deposit number is CCTCC NO: V201861. The attenuated strain of type 3 duck hepatitis A virus is safe to ducklings, has no pathogenicity, can protect against homologous strong virus challenge, and has practical and extensive application value.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, in particular to a type 3 duck hepatitis A virus attenuated strain CH-P60 and application thereof. Background technique [0002] Duck virus hepatitis (DVH) is an acute and highly contagious disease caused by ducklings infected by duck hepatitis virus (DHV). At present, the disease exists in major duck raising areas in the world, and has the characteristics of intermittent outbreaks and endemicity. It is one of the main diseases that endanger the duck industry. The disease mainly affects ducklings less than one month old. It has the characteristics of acute onset, rapid spread, short course of disease and high mortality. "; The pathological changes were mainly liver enlargement and inflammation and a large number of hemorrhagic spots at autopsy. The disease is mainly caused by Duck hepatitis Avirus (DHAV) belonging to the genus Avihepatovirus of the family Picornaviridae. DHAV has three ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12N7/08A61K39/125A61P31/14
Inventor 程安春文兴建汪铭书
Owner SICHUAN AGRI UNIV
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