Method for preparing TW1 type avian infectious bronchitis virus positive serum from SPF chickens
A chicken infectious and positive serum technology, applied in the direction of chemical instruments and methods, antiviral immunoglobulin, immunoglobulin from serum, etc., can solve complex and variable biological characteristics, pathogenicity and tissue tropism, cross Protection and serum neutralization efficiency are poor, and disease diagnosis and prevention work are difficult, so as to achieve the effect of improving the level of prevention and control, controllable quality, and stable source
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0039] Embodiment 1——Virus content and purity detection of virus species for immunization
[0040] Carry out virus content determination to virus species according to the following method, AH1103 chicken embryo allantoic fluid is made 10 times of serial dilutions with sterile saline, take 10 -5 、10 –6 、10 –7 、10 –8 For 4 dilutions, 5 10-day-old SPF chicken embryos were inoculated into each allantoic cavity, 0.1ml per embryo, and incubated at 36-37°C. Chicken embryos that died before 24 hours were discarded, and chicken embryos that died within 24 to 144 hours were taken out at any time, until 144 hours, all live embryos were taken out. The dead chicken embryos and surviving chicken embryos in 24-144 hours after inoculation have specific lesions such as dehydration, curling up, and small development (the weight of the inoculated fetus is more than 2g lower than that of the lightest fetus in the control) and other specific lesions are judged as infection, and the EID is calc...
Embodiment 2
[0045] Embodiment 2——Genotyping of chicken infectious bronchitis virus IBV CK / CH / XNGD / 140 strains
[0046] According to the sequence of chicken infectious bronchitis virus (IBV) S1 gene included in GenBank, primers (sequence 1 and sequence 2) were designed for the conserved region after alignment, and were synthesized by Shanghai Sangon Bioengineering Co., Ltd. The primer sequences are as follows:
[0047] Sequence 1 upstream primer S1–F: 5′–atgttgggga agtcactgtt–3′20
[0048] Sequence 2 downstream primer S1–R: 5′–acgtctaaaa cgacggctt–3′19.
[0049] IBV CK / CH / XNGD / 140 strains were inoculated into 10-day-old SPF chicken embryos, and the allantoic fluid of 36-42h chicken embryos was collected. The S1 gene of CK / CH / XNGD / 140 strain was amplified by RT-PCR, and the amplified product was cloned, sequenced, and compared with the gene sequence. Viral RNA was extracted from allantoic fluid according to the instructions of the RNA extraction kit of Tiangen Biochemical Technology (Bei...
Embodiment 3
[0057] Embodiment 3——Preparation of chicken infectious bronchitis virus IBV CK / CH / XNGD / 140 strain inactivated vaccine
[0058] Inoculate 10-day-old SPF chicken embryos with CK / CH / XNGD / 140 virus seeds, and collect the allantoic fluid of 36-42h chicken embryos. The harvested chicken embryo virus liquid was centrifuged at 8000 rpm for 10 min, and the supernatant was harvested. The centrifuged virus fluid was dialyzed and concentrated to 1 / 5 of the original volume, and inactivated with formaldehyde at a final concentration of 0.15% at 37° C. for 20 h. Preparation of oil phase: heat white oil (94 parts) to 80°C, then add Siben-80 (6 parts), mix evenly, heat to 121°C for 30 minutes, cool for later use. Preparation of aqueous phase: add sterilized Tween-80 (4 parts) to inactivated antigen solution (96 parts), and mix well. Emulsification: Measure the water phase which is 1 / 3 of the volume of the oil phase, add it to the oil phase, and mix at 1500r / min for 2min. Adjust the speed to...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com