Eurasian Avian H1N1 Subtype Swine Influenza Cold Adapted Attenuated Strain and Its Application

A technology for H1N1 and swine flu, applied in veterinary vaccines, antisense single-stranded RNA viruses, medical preparations containing active ingredients, etc., can solve the problems of lack of virus cross-protection, long residual time, etc.

Active Publication Date: 2020-08-04
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Generally speaking, inactivated vaccines have good immune protection and can protect against the same subtype of influenza virus. The disadvantage is that they cannot induce effective mucosal immune antibodies and cellular immune responses. Therefore, they lack protection against other subtypes or lineage viruses. Cross-protection, and the route of vaccination is usually through intramuscular injection, the vaccine needs to be added with corresponding adjuvants, and there will be a long residual time in the body

Method used

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  • Eurasian Avian H1N1 Subtype Swine Influenza Cold Adapted Attenuated Strain and Its Application
  • Eurasian Avian H1N1 Subtype Swine Influenza Cold Adapted Attenuated Strain and Its Application
  • Eurasian Avian H1N1 Subtype Swine Influenza Cold Adapted Attenuated Strain and Its Application

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Example 1 Preparation of Eurasian Avian H1N1 Subtype Swine Influenza Cold-adapted Attenuated Vaccine Strain A / swine / Guangxi / 18 / 2011ca (H1N1)

[0027] 1. Take out 1 part of the Eurasian avian type H1N1 subtype swine influenza virus strain A / swine / Guangxi / 18 / 2011 (H1N1) (abbreviated as GX18) preserved in our laboratory, put it on ice after dissolving, and measure hemagglutination ( HA) value, and calculate 4HA units / 0.2mL;

[0028] 2. The allantoic cavity of 10-day-old SPF chicken embryos was inoculated with 0.2 mL of GX18 virus dilution (diluted in pH 7.2-7.4 phosphate buffer solution) containing 4HA units;

[0029] 3. Cultivate at 33°C for 72-96 hours, measure the HA value, and when the HA value reaches 128-256, harvest the virus liquid and use it as the seed virus for the next incubation temperature (in units of 1°C, the gradient decreases), and continue to step 2 experiment.

[0030] In this way, 10-day-old SPF chicken embryos were uploaded for 17 generations, and a...

Embodiment 2

[0035] Example 2 Cold-adapted (cold-adapted, ca) determination of swine influenza cold-adapted attenuated vaccine strain GX18ca

[0036] Cold adaptability (ca) is to compare the replication titer of virus strains on chicken embryos under the suitable growth temperature condition of 33°C and the low temperature growth condition of 25°C. A virus is considered cold-adapted if its growth titer on chicken embryos at 25°C is within 100-fold of its growth titer at 33°C, and vice versa .

[0037] Method: Dilute the virus GX18ca and the corresponding wild virus GX18 by 10 times on ice, that is, 100 μL virus + 900 μL PBS, mix well, pipette 100 μL virus dilution into the next tube of 900 μL PBS, mix well, and then And so on, all the way down to 10 -9 . The diluted virus was inoculated into four 10-day-old SPF chicken embryos for each dilution, and the inoculated chicken embryos were placed in a low-temperature incubator at 25°C for incubation for 72 hours. And inoculate another 4 10-...

Embodiment 3

[0040] Example 3 Determination of temperature sensitivity (temperaturesensitive, ts) of swine influenza cold-adapted attenuated vaccine strain GX18ca

[0041] Temperature sensitivity (ts) is to compare the replication titer of virus strains on chicken embryos under the conditions of suitable growth temperature of 33°C and high temperature of 39°C. If the growth titer of the virus on chicken embryos at 39°C is more than 100 times lower than that at the appropriate growth temperature of 33°C, then the virus is considered to be temperature-sensitive, and vice versa Of course.

[0042] Method: serially dilute virus 10 2～7 times, determine the EID of GX18ca and the corresponding wild virus GX18 at 33°C or 39°C 50 , after incubation for 72h, the virus replication situation at each temperature was detected by hemagglutination test, and the half chicken embryo infectious dose (EID) was calculated by the Reed-Muench method. 50 ). The replication of GX18 wild virus at 33°C and 39°C ...

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Abstract

The invention discloses a Eurasian avian like H1N1 swine influenza A virus cold-adapted attenuated virus strain and an application thereof. Eurasian avian like H1N1 swine influenza A viruses A / swine / Guangxi / 18 / 2011 (H1N1) are subjected to serial sub-culturing at the temperature of 33-25 DEG C to obtain the Eurasian avian like H1N1 swine influenza A virus cold-adapted attenuated virus strain named as a swine influenza cold-adapted attenuated virus strain A / swine / Guangxi / 18 / 2011ca (H1N1), the strain is collected in a China Center for Type Culture Collection, and the collection number of the strain is CCTCC NO. V201720. Researches show that an attenuated live vaccine prepared from the cold-adapted attenuated virus strain can resist attack of swine influenza A virus wild strains mainly epidemic in Chinese swine herds, and is of great significance for controlling prevalence and human infection of Eurasian avian like H1N1 and H1N2 swine influenza A viruses.

Description

technical field [0001] The present invention relates to a strain of influenza virus attenuated vaccine strain, in particular to a kind of Eurasian avian (Eurasian avian like, EA) H1N1 subtype swine influenza virus (Swine influenza A virus, SIV) gradually reduced in SPF chicken embryo The broad-spectrum cold-adapted attenuated vaccine strain obtained by temperature cultivation also relates to the application of the vaccine strain in preventing and controlling EAH1N1 subtype swine influenza virus. The invention belongs to the technical field of veterinary drugs. Background technique [0002] Swine influenza is an acute and contagious respiratory infectious disease of pigs caused by swine influenza A virus (SIV). It is characterized by sudden onset, cough, dyspnea, fever and rapid regression. Usually, swine flu has low mortality rate and high morbidity rate. It is widely prevalent in the world and has caused huge economic losses to the pig industry. [0003] There are three ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K39/145A61P31/16
CPCA61K39/12A61K2039/5254A61K2039/552C12N7/00C12N2760/16134
Inventor 杨焕良陈化兰陈艳乔传玲
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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