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Pigeon paramyxovirus 1 strain AF-1 and application thereof

An AF-1, paramyxovirus technology, applied in the direction of viruses, antiviral agents, virus antigen components, etc., can solve the problems of unsatisfactory protection effect and genotype differences, and achieve good safety, good immunogenicity, and multiplication droplets. high degree of effect

Inactive Publication Date: 2016-09-14
武汉智健动物保健有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although pigeon paramyxovirus type 1 belongs to the same paramyxovirus as chicken Newcastle disease virus, the protective effect is not satisfactory due to the differences in their genotypes

Method used

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  • Pigeon paramyxovirus 1 strain AF-1 and application thereof
  • Pigeon paramyxovirus 1 strain AF-1 and application thereof
  • Pigeon paramyxovirus 1 strain AF-1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The isolation and identification of embodiment 1 virus

[0023] The visceral tissues of dead pigeons were collected from a pigeon farm in Anhui, and after RNA extraction and detection by RT-PCR, it was determined that they were pigeon paramyxovirus type 1, and the virus strain was named as pigeon paramyxovirus PPMV-1AF-1 strain. The specific operation steps are as follows:

[0024] 1. Chicken embryo multiplication

[0025] Add double antibody (final concentration: 10000U / ml) to the disease material treatment solution, incubate at 33-35°C for 30 minutes, inoculate 9-11-day-old SPF chicken embryos through the chorioallantoic cavity, and inoculate 4 chicken embryos per disease material 0.2ml per embryo. The inoculated chicken embryos were incubated at 33-35°C for 96 hours, the humidity was maintained at 60-65%, and the eggs were illuminated regularly. Chicken embryos that died within 24 hours were discarded. Take the dead and surviving chicken embryos that have been de...

Embodiment 2

[0045]The preparation of embodiment 2 inactivated vaccine 1

[0046] 1. Determination of virus content

[0047] The virus liquid was diluted serially 10 times with sterilized physiological saline, and 10 -6 、10 -7 、10 -8 Inoculate 10-day-old SPF chicken embryos into the allantoic cavity of 3 dilutions, inoculate 5 eggs for each dilution, 0.1ml / embryo, record the infection status of chicken embryos in 24-120 hours, and calculate EID 50 , the virus content per 0.1ml should be ≥10 7.0 EID 50 . After determination, the EID after harvesting the allantoic fluid was mixed 50 See the table below:

[0048] Table 1 Reed-Muench method to calculate EID 50 result

[0049]

[0050] The calculation method is as follows:

[0051]

[0052] wxya 50 = logarithm of highest dilution of virus above 50% infection + distance ratio x difference between dilutions.

[0053] Distance Scale = 0.78

[0054] wxya 50 =-7+(-1×0.78)=-7.78

[0055] EID 50 =10 -7.78 / 0.1ml

[0056] 2. Vir...

Embodiment 3

[0068] 1. Vaccine safety test

[0069] With 15 paramyxovirus type I antibody-negative pigeons at the age of 21 to 30 days, 10 of them were subcutaneously injected with 1 ml of vaccine in each neck, and the other 5 were used as controls. After 14 days of observation, there should be no local and systemic adverse reactions caused by the injection of the vaccine. The test results are shown in the table:

[0070] Table 3 Safety detection of paramyxovirus type I antibody-negative pigeons in 21-30 days old pigeons inoculated with overdose of inactivated vaccine

[0071] Before vaccination after inoculation immune group 10 / 10 normal 10 / 10 normal control 5 / 5 OK 5 / 5 OK

[0072] 2. Immunity and virus attack protection experiment

[0073] 2.1. Immunity

[0074] With 15 paramyxovirus type I antibody-negative pigeons at the age of 21 to 30 days, 10 pigeons were subcutaneously injected with 200 μl of vaccine in each neck, and the other 5 were used as ...

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Abstract

The invention discloses a pigeon paramyxovirus 1 strain AF-1 and an application thereof. The strain is a pigeon paramyxovirus 1 strain AF-1, and the preservation number of the strain is CCTCC NO:V 201613. A method for preparing an inactivated vaccine through using the pigeon paramyxovirus 1 strain AF-1 comprises the following steps: culturing the pigeon paramyxovirus 1 strain AF-1 to obtain an AF-1 strain virus solution, and adding beta-propiolactone to the AF-1 strain virus solution; inactivating the obtained virus solution to obtain a mixed solution; adding tween-80 to obtain an aqueous solution; adding aluminum stearate to white oil while stirring until the obtained oil is completely transparent, and adding tween-80 to obtain an oil solution; and slowly adding the aqueous solution to the oil solution, placing the obtained solution mixture in an emulsifying tank, and emulsifying the solution mixture to obtain the inactivated vaccine. The inactivated vaccine helps pigeons to effectively resist infection induced by pigeon paramyxovirus 1, greatly reduces the loss, promotes the development of the pigeon industry, is safe and effective, and can be applied to production practices.

Description

technical field [0001] The invention relates to pigeon type 1 paramyxovirus, in particular to a pigeon type 1 paramyxovirus strain AF-1 and application thereof. Background technique [0002] Pigeon plague, also known as pigeon Newcastle disease or paramyxovirus disease, is caused by pigeon paramyxovirus type 1 (Pigeon Paramyxovirus 1; PPMV-1). The disease is widespread and mainly causes pigeon diarrhea, neurological symptoms and death, and causes great obstacles to the healthy development of the global pigeon industry. Pigeons of different breeds and ages are susceptible. Generally speaking, the incidence rate of pigeon can be as high as 80~90%. According to the specific immune status and feeding conditions, the mortality rate is roughly 30-80%. [0003] Because pigeons do not directly affect the daily life of most people, and the amount of breeding is relatively small. Therefore, it has not aroused the awareness of relevant industries and the attention of the country fo...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/17A61P31/14
CPCC12N7/00A61K39/12A61K2039/5252A61K2039/552A61K2039/55511C12N2760/18121C12N2760/18134
Inventor 陈博
Owner 武汉智健动物保健有限公司
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