GB/T 14926.23-2001 Detection method of Sendai virus in experimental animals

This standard specifies the detection method and reagents for Sendai virus (SV). This standard is applicable to the detection of Sendai virus in mice, rats, guinea pigs, hamsters and rabbits. GB/T 14926.23-2001 Detection method of Sendai virus in experimental animals GB/T14926.23-2001 Standard download decompression password: www.bzxz.net

ICS65.020.30
National Standard of the People's Republic of China
GB/T 14926.23-2001
Laboratory animal
Microbiological examination methods (4)
Laboratory animalMicrobiological examination methods2001-08-29Promulgated
General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China
Implementation on 2002-05-01
GB/T14926.23—2001
This standard is a revision of GB/T14926-23-—19944 "External fluorescence test method for laboratory animals, Sendai virus test method", with the addition of the detection of virus antibodiesThis standard is authorized and supervised by the Ministry of Science and Technology of the People's Republic of China, and the drafting unit of this standard is the Chinese Society of Animal Science. This standard was mainly drafted by Da He Shanming. This standard was first issued in January 1994. National Standard of the People's Republic of China Laboratory Animals Sendal virus detection method Laboratory animal Method for esamination of Sendal virus (SV) This standard specifies the detection method and reagents of Sendal virus (SV). This standard is applicable to the detection of Sendal virus in Tibetans, Tibetans, guinea pigs, hamsters and rats. Reference Standards GB/T14926-23-2001 GB/T14926.23-1994 The provisions contained in the following standards constitute the provisions of this standard through reference in this standard. When this standard is published, the versions shown are valid. All standards will be revised. All parties using this standard should discuss the possibility of using the latest version of the following standards. GB/T14926.50-2001
GB/T14926.51—2001
GB/T 14926.52-2001
GB/T14926.54—2001
Laboratory animals
Laboratory animals
Laboratory animalsbzxZ.net
Laboratory animals
Enzyme-linked immunosorbent assay
Immunotoxicity test
Immunofluorescence test
Hemoglobin inhibition test
According to the principle of immunology, SV antigen is used to detect Sendai virus antibodies in the sera of mice and rats: or according to the principle that SV can absorb erythrocytes of chickens and rats under certain conditions, and this ability of erythrocytes can be inhibited by specific antibodies , detect Sendai virus antibodies in mouse, rat, guinea pig, and hamster immune sera. Main reagents and equipment
4.1 Reagents:
4.1-1 ELISA antigen
4.1.1-1 Specific antigen
Infect 9d SPF chickens with SV and press the allantoic cavity. Culture in a 36℃ incubator. After 72h, freeze at 4℃+ and collect the allantoic fluid aseptically the next day. Centrifuge at 4℃2000r/min for 10min. Use 0.5% chicken or guinea pig red blood cells and SV positive serum to do hemagglutination and hemagglutination inhibition tests to verify its virus specificity and hemagglutination titer. The supernatant is then concentrated by ultracentrifugation to make ELISA antigen. 41.1-2 Normal antigen
9-day-old SPF chicken urine fluid.
4.1.2 Antigen slice
SV infected BHK21 cells, 2~3 days after inoculation, when the lesions reached ++~+++, use lysate to digest and disperse, wash with PBS, and wash the slice. After drying at room temperature, fix with cold acetone for 10min-20c and store. Approved by the General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China on 2001-08-29 and implemented on 2002-05-01
4.1-3 Hematin
See Preparation of ELISA specific antigens,
4.1.4 Positive serum
GB/T 14926-232001
SV antigen antiserum obtained from clean or SPF mice, rats, rats, urine or normal grade rabbits, 4.1.5 negative serum
clean or SPF rats, rats, guinea pigs, blood and blood without diseases 4.1-6 enzyme conjugate
horseradish peroxide labeled single or rabbit anti-mouse, rat, guinea pig, hamster IgG antibody, used to detect corresponding animal serum antibodies, peroxide labeled sheep anti-rabbit IgG antibody, used to detect rabbit serum antibodies, horseradish peroxide labeled coccal protein A (SPA can be used for mouse, hamster, rabbit blood antibody examination 4.1.7 fluorescein isothiocyanate labeled sheep or rabbit anti-mouse, rat, hamster IgG, used to detect corresponding animal serum antibodies, This is used to detect the antibody
Isocyanate fluorescein labeled goat anti-rabbit IgG plate
4.2 Equipment
4-21 Label avoidance instrument
4.2-2 Fluorescence microscope,
4-2-3 Ordinary microscope.
4-2.437℃ incubator or
5 Detection method
5.1 Use ELISAGB/T14926
5 2 Use IFA method
5.3 Use IEA method
5-4 Use HAT method
6 Nesting judgment
For positive test results
B/T14926.5
B/T14926.51—2001), perform electrochemical examination. 7114926-
a method or
according to the judgment result, a method type
is judged as positive
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