This standard specifies the detection method and reagents for pneumonia virus of mice (PVM). This standard is applicable to the detection of PVM in mice, rats, hamsters and guinea pigs. GB/T 14926.24-2001 Detection method for pneumonia virus of mice in experimental animals GB/T14926.24-2001 Standard download decompression password: www.bzxz.net

ICS65.020.30
National Standard of the People's Republic of China
GB/T 14926.24-2001
Laboratory Animals
Microbiological Examination Methods (4)
Laboratory AnimalsMicrobiological Examination Methods2001-08-29Promulgated
General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China
Implemented on 2002-05-01
GB/T14926.24—2001
Test Methods for Pneumonia Virus in Mice" is revised, except for hemagglutination inhibition. This standard is a revision of GB/T14926.24-1994% Laboratory Animal Test Methods, and some texts have been modified. This standard is proposed and managed by the Ministry of Science and Technology of the People's Republic of China. The chapter unit of this standard is the Chinese Society of Laboratory Animal Science. This standard is intended for the enjoyment of the public and is used for the first time in January 1994.
National Standard of the People's Republic of China
Laboratory animal-Method for examination of pneumonia virus of mice (PVM) This standard specifies the method and reagents for the examination of pneumonia virus of mice (PVM). This standard is applicable to the examination of PVM in mice, rats, runners and glandular rats. Referenced Standards
GB/T14926-24—2001
GB/T14926-24—1994
The provisions contained in the following standards, which are cited in this standard, constitute the provisions of this standard. When this standard is published, the versions are valid and all standards will be subscribed. All parties using this standard should explore the possibility of using the latest version of the following standards. Enzyme-linked immunosorbent assay
GB /T14926-50—2001
Laboratory animals
GB/T14926.51—2001
GB/T14926.52—2001
Laboratory animalsbZxz.net
Laboratory animals
Immunoenzyme test
Immunofluorescence test
According to the principles of immunology, PVM antigen is used to detect PVM antibodies in the serum of mice, rats, hamsters and guinea pigs. Main reagents and equipment
4-1 Reagents:
4.1.1 ELISA antigen
4.1.1-1 Specific antigen
After the onset of the disease, take the lungs of PVM-infected mice, grind them into a 10% suspension, centrifuge them at 3000r/min for 10min, take the supernatant and infect BHK21 cells, adsorb for 1.5~2h+ culture in maintenance medium for 10~14d, harvest when the cytopathic effect reaches +++, freeze-thaw three times or treat with ultrasound, remove cell debris by low-speed centrifugation, and then make the supernatant into ELISA antigen after centrifugation at a certain speed. 4.1.1.2 Normal antigen
BHK21 cells are cryo-broken and broken with acid, and the cell debris are removed by low-speed centrifugation to obtain the supernatant. 4.1.2 Antigen slice
PVM infected BHK21 cells. Cultured for 5~7 days. When the lesions reached ++~+++, the cells were dispersed by trypsin digestion, washed with PBS, sliced, dried at room temperature, and then cold-fixed for 10 minutes and stored at -20°C. 4.1.3 Positive serum
PVM immune serum or antiserum obtained from SPF mice, rats, guinea pigs, and hamsters. 4.1.4 Positive serum
Approved by the General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China on August 29, 2001 and implemented on May 1, 2002
SPF mouse, rat, guinea pig, and hamster serum. 4.1.5 Enzyme conjugates
GB/T14926-24—2001
Horseradish peroxide alcohol is used to label single or rabbit anti-mouse, rat, guinea pig, hamster IgG antibodies, and the corresponding animal serum antibodies are detected by hand. Horseradish peroxide-labeled Staphylococcus protein A (SPA) can be used to check the serum antibodies of mice, rats, and hamsters. 4.1.6 Fluorescein is used to label sheep or rabbit anti-mouse, rat, guinea pig, hamster IgG antibodies, and the corresponding animal serum antibodies are detected by hand. 4.2 Equipment
42.1 Only required
4.2.2 Fluorescence microscope.
4.2.3 Fluorescence microscope.
4.2.437C incubator or water cream
5 Detection method
Use ELISA method (see GB/T14926.502001) for serological detection. 51
Use IFA method (see GB/T14926.52-2001) for serological detection. 52
Use IEA method (see GB/T14926-51-2001) for serological detection. Results judgment
For positive test results, use the same method or another method to retry. If it is still positive, it is considered positive. Result report
Make a report based on the judgment result.
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