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Avian infectious bronchitis virus IBYZ and application thereof

A chicken infectious and bronchitis technology, applied in the field of virology, can solve the problems of difficulty in evaluating the immune protection effect of IB vaccine, lack of pathogenesis models of highly pathogenic strains, and large differences in pathogenicity

Inactive Publication Date: 2020-03-17
JIANGSU INST OF POULTRY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the QX genotype strains that are mainly prevalent in chicken flocks in my country are clinically manifested as pathogenic to kidney tissue, and can cause different degrees of death in infected chickens, but the pathogenicity differences between different isolates are relatively large. Large and lack of highly pathogenic strains to establish a unified pathogenesis model, which has caused difficulties for further research on the pathogenic mechanism of the virus and the evaluation of the immune protection effect of the IB vaccine

Method used

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  • Avian infectious bronchitis virus IBYZ and application thereof
  • Avian infectious bronchitis virus IBYZ and application thereof
  • Avian infectious bronchitis virus IBYZ and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Isolation and Identification of Chicken Infectious Bronchitis Virus

[0033] 1. Virus isolation

[0034] In 2011, a strain of chicken infectious bronchitis virus was isolated from dead chickens in a large-scale farm in Jiangsu Province. The specific method is: aseptically collect the kidney tissue of sick and dead chickens, cut it into pieces and grind it, add penicillin and streptomycin in sterilized saline with a content of 1000 IU / mL each according to 1:5 (v / v), freeze it repeatedly after homogenizing Melt twice, centrifuge at 5000r / min for 5min, take the supernatant and filter to sterilize, inoculate 4 11-day-old SPF chicken embryos through the allantoic cavity, 0.2mL / piece, and incubate at 37°C for 48h; discard dead chicken embryos within 24h , collect 24h ~ 48h chicken embryo allantoic fluid as seed poison, and conduct continuous passage through SPF chicken embryos.

[0035] 2. Dwarfing test of chick embryos

[0036] The P6 seed virus of the IBYZ strain was dil...

Embodiment 2

[0042] Whole Genome Sequencing of Chicken Infectious Bronchitis Virus Isolates

[0043] 1. Extraction and reverse transcription of viral genome RNA

[0044] Using RNAiso Plus (TaKaRa), the genomic RNA of the isolate was extracted according to the steps in the kit instruction manual. Using the extracted viral genomic RNA as a template, reverse transcription was performed using PrimeScript 1st Strand cDNASynthesis Kit (TaKaRa) according to the instruction manual.

[0045] 2. Segmented amplification and sequence determination of the genome

[0046] According to the IBV whole genome sequence NC001451 (Beaudette) and FJ807652 (H120) published on the International GenBank (GenBank) after multiple comparisons, the whole genome sequencing primers of the IBYZ strain were designed in relatively conservative regions (see Table 1). Synthesized by Shanghai Sangon Bioengineering Co., Ltd. Using the reverse transcription product of the above-mentioned virus genome as a template, the genom...

Embodiment 3

[0056] Animal regression experiment of chicken infectious bronchitis virus isolate

[0057] Take fresh allantoin from IBYZ isolate and dilute to EID with PBS 50 for 10 5.0 / 0.1mL, inoculate 10 1-day-old (or 7-day-old) SPF chicks, 0.1mL / piece, observe and record the morbidity and mortality of the chicks after inoculation, and set up a negative control group at the same time. On the 3rd day after inoculation with IBYZ, the chicks began to show obvious clinical symptoms, mainly manifested as depression, mouth breathing, runny nose, head shaking and other symptoms. Deaths began on day 3 or 4. The dead chickens were autopsyed, and the main pathological changes were enlarged and pale kidneys, dilated oviducts and renal tubules, and a large amount of urate deposited. image 3 ). The morbidity rate of 1-day-old SPF chicks was 100% (10 / 10), and the mortality rate was 60%; the morbidity rate of 7-day-old SPF chicks was 100% (10 / 10), and the mortality rate was 50%. The strain can es...

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Abstract

The invention provides an avian infectious bronchitis virus IBYZ and an application thereof, and relates to the technical field of virology. The preservation number of the avian infectious bronchitisvirus IBYZ is CGMCC No. 14682. The avian infectious bronchitis virus IBYZ has typical coronavirus genome coding structure characteristics, and belongs to a predominant prevalent QX genotype at present. The strain can cause typical clinical symptoms of SPF chicks, the death rate of seven-day-old SPF chicks is 50% or above, a stable pathogenesis model can be established through the strain, and the strain can be used for researching the pathogenesis of the virus and evaluating the immune protection effect of IB vaccines.

Description

technical field [0001] The invention belongs to the technical field of virology, and in particular relates to a strain of chicken infectious bronchitis virus IBYZ and its application. Background technique [0002] Chicken infectious bronchitis (Infectious bronchitis, IB) is one of the second-class infectious diseases of poultry stipulated by the International Veterinary Bureau. It is an acute and highly contagious infection caused by infectious bronchitis virus (Infectious bronchitis virus, IBV). sick. Because IBV is very easy to mutate, new mutant strains appear constantly, and the cross protection between different strains is weak, so that the disease often breaks out in immune and non-immune poultry flocks, causing huge economic losses and seriously restricting the The healthy development of my country's poultry industry. [0003] IBV belongs to the Gamma coronavirus of the Coronaviridae family. It is a single-stranded positive-sense RNA virus with an envelope and no seg...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/215A61P31/14A01K67/027
CPCA01K67/027A01K2267/0337A61K39/12A61P31/14C12N7/00C12N2770/20021C12N2770/20034
Inventor 周生姜逸俞燕程旭赵秀美唐梦君高明艳韦玉勇
Owner JIANGSU INST OF POULTRY SCI
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