This standard specifies the detection method and reagents for mousepox virus (Ect). This standard is applicable to the detection of mouse Ect. GB/T 14926.20-2001 Detection method of mousepox virus in experimental animals GB/T14926.20-2001 Standard download decompression password: www.bzxz.net

ICS65.020.30
National Standard of the People's Republic of China
GB/T14926.20-2001
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/T14926-20-2001
"Test Method for Mouse Pox Virus in Laboratory Animals" is revised. The "Test Method for Mouse Pox Virus in Laboratory Animals" is renamed "Test Method for Mouse Pox Virus in Laboratory Animals" to improve the sensitivity of the test method. Vaccinia virus is no longer used as a test method for mouse pox virus. Alternative antigens for poxvirus antibody detection. In addition to the virus detection related content specified in 5.2 of GB/T14926-20-1994, the immunoenzyme histochemistry method is added as a detection method for virus antigens. This standard is proposed and coordinated by the Ministry of Science and Technology of the People's Republic of China. The drafting unit of this standard is: Chinese Society of Laboratory Animal Science. The main drafter of this standard is He Zhengming.
This standard was first issued in January 1994.
National Standard of the People's Republic of China
Laboratory Animals
Euctoxvirus Detection Method
Laboratory animalMethod for examination ofEetromelia wirus (Eet..)
This standard specifies the detection method and reagents of Eet. This standard is applicable to the detection of mouse Ect.
Cited Standards
GB/T14926.202001
Representative GB/T14826:20——1994
The provisions contained in the following standards, when cited in this standard, constitute the provisions of this standard. When this standard is published, the versions shown are valid. All standards will be revised, and the parties using this standard should ensure the possibility of using the latest versions of the following standards. GB/T14926.50—2001
GB/T 14926.51—2001
GB/T14926-52—2001
GB/T 14926- 552001
In vivo animals
Experimental animals
Experimental animals
Experimental animals
Enzyme-linked immunosorbent assay
Immunoenzyme assay
Immunofluorescence assay
Immunoenzyme histochemistry
According to the principle of immunology, Ect antigen is used to detect Ect antibody in mouse serum: or known Ect antibody is used to detect Ecr antigen in rat tissue
4 Main reagents and equipment
4.1 Reagents
4.1.1 ELISA antigen
4.1.1.1 Specific antigen
Use Ect. to detect BHK2I cells. When the lesions reach 10 to 100 mm, they are harvested. After freeze-thaw three times or ultrasonic reduction, the cell debris is removed by low-speed centrifugation, and the supernatant is further shrunk by centrifugation to make ELISA antigen. 4.1.1.2 Normal Antigen
BHK21 cells were frozen and thawed, and the supernatant obtained by low-concentration centrifugation to remove the positive plate 4.1-2 Antigen Plate
Ect-infected BHK21 cells were inoculated for 2~3 days. When the lesions reached ++~+++, they were digested and dispersed with pancreatic alcohol, washed and plated with PBS, dried at room temperature, fixed with cold acetone for 10 minutes, and stored at -20°C. 4.1.3 Positive Serum
Ect antigen was inactivated with β-propiolactone, and the antiserum was obtained by immunizing clear or SPF mice. 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
4.1.4 Positive Serum
Clean or SPF mouse serum.
4.1.5 Enzyme conjugate
GB/T14926-202001
Sheep or rabbit anti-mouse IgG antibody labeled with peroxidase: or Staphylococcus aureus protein A (SPA) labeled with isothiocyanate fluorescein, or rabbit anti-mouse IgG antibody.
42-1 Alcohol labeling instrument.
4.2-2 Fluorescence microscope
42.3 Ordinary microscope.
Five-stage slide machine or frozen slide machinebzxZ.net
4.2.537C Cultivation box or water box.
5 Detection method
Use ELISA method (see GB/T14926.50-2001) for serological testing 5.1
Use IFA method (see GB/T14926.52-2001) for serological testing. Use IEA method (see GB/T14926.51-2001) for serological testing. 5.3
Use immunoenzyme plastic chemistry method (see GB/T14926.55-2001) for sperm antibody testing. Results
For the test results, use the same method or another method to retry. If it is still negative, it is judged as positive. Result report
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