This standard specifies the detection methods and reagents for Hantavirus (HV). This standard is applicable to the detection of HV in mice and rats. GB/T 14926.19-2001 Detection methods for Hantavirus in experimental animals GB/T14926.19-2001 Standard download decompression password: www.bzxz.net

ICS65.020.30
National Standard of the People's Republic of China
GB/T14926.19-2001
Laboratory Animals
Microbiological Examination Methods (4)
Laboratory Animal Microbiological Examination Methods2001-08-29Promulgated
People's Republic of China
General Administration of Quality Supervision, Inspection and Quarantine
2002-05-01Implementation
GB/T14926.192001
This standard is a revision of GB/T14926-19—1994 Method for Detection of Epidemic Hemorrhagic Fever Viruses in Laboratory Animals. The "Method for Detection of Epidemic Hemorrhagic Fever Viruses in Laboratory Animals" is renamed as "Method for Detection of Hantaviruses in Laboratory Animals". The contents of "5. Detection methods and results" in GB/T 14926.15 of 1994 have been removed, and the static immunosorbent test and immunofluorescence test for detecting virus antibodies have been added. This standard is proposed and managed by the Ministry of Science and Technology of the People's Republic of China. The origin of the standard is China Laboratory Animal Society. The main drafter of this standard is: Mao Zhengming. This standard was first issued in January 1994. 1 National Standard of the People's Republic of China Laboratory animal-Method for exsmination ofHantavirus This standard specifies the detection methods and reagents of Hantan virus (CHV). This standard is applicable to the detection of CHV in rats and mice. The referenced standard GB/T14926.19-2001 represents GB/T14926.19-1994. 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, and the parties using this standard should explore the possibility of using the latest versions of the following standards. GB/T14926-50-2001
Laboratory animals
GB/T14926.52-2001||tt| |3 Principle
Experimental animals
ELISA
Immunofluorescence test
According to the principle of immunology, HIV antigen is used to detect HIV antibodies in the serum of mice and rats. 4 Main reagents and equipment
4.1 Reagents
4.1.1 ELISA antigens
4.1.1.1 Specific antigens
In a biosafety cabinet, E6 cells infected with Hantaan or Seoul strains are used. When the specific fluorescence reaches +++, the culture can be harvested. After freeze-thaw or ultrasonic treatment, low-speed centrifugation is used to remove cell debris, and the supernatant is concentrated by ultracentrifugation to prepare ELISA antibodies.
4-1.1-2 Positive Common antigensWww.bzxZ.net
After E6 tobacco fat is acid-crushed, the supernatant obtained by low-speed centrifugation to remove the cell fragments, 4.12 Antigen slices
In a biological safety cabinet, infect E6 cells with Hantan or Seoul strains, replace the maintenance medium every 2-3 days, culture for 7-10 days, and measure the specific fluorescence in the cells by IFA. When the fluorescence reaches ++10, the cells are dispersed with trypsin, washed with PBS, smeared, and irradiated at 20cm under ultraviolet light for 30 minutes while operating at room temperature, fixed with cold acetone for 10 minutes and stored at +-20C, 4.1.3 Antiserum
Use β-propiolactone flexible HV antigen, immune serum or antiserum obtained from SPF mice or rats. 4-1.4||tt| |Invasive serum
The General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China approved on August 29, 2001, and sold on May 1, 2002. Specification
Serum of SPF mice or rats,
41.5 Binding substances
GB/T14926-192001
Horseradish peroxidase-labeled goat or rabbit anti-mouse, rat IgG antibodies, used to detect corresponding animal serum antibodies: horseradish peroxidase-labeled Staphylococcus aureus protein A (SPA>, used to detect mouse serum antibodies, 4.1.6 Fluorescein isothiocyanate-labeled goat or rabbit anti-mouse, rat IgG antibodies, used to detect corresponding animal serum antibodies 4-2 Equipment
4.2.1 Alcohol labeling instrument.
4.2.2 Fluorescence microscope,
4.2.337C culture box or water box.
Test method
5-1 Using ELISA method C
52 Using IFA method
For positive test results
Result report
Based on the determination
One of the methods
is judged as positive
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