GB/T 19180-2020 in English
VALIDDiagnostic techniques for bovine spongiform encephalopathy
- Issued on:2020-12-14
- Implemented on:2020-12-14
- File Format:PDF
- Delivery:Via email within 1~3 business days
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《GB/T 19180-2020牛海绵状脑病诊断技术》由TC181（全国动物卫生标准化技术委员会）归口，主管部门为农业农村部。
Introduction
Background of Standard Revision and Technological Evolution
GB/T19180-2020, as the new standard replacing the 2003 version, reflects the three core advances in mad cow disease diagnostic technology:
| Technical Dimensions | 2003 Version | 2020 Version |
|---|---|---|
| Diagnostic Methods | Only clinical symptoms + histopathology + immunohistochemistry | Newly added Western blotting as a diagnosis method |
| Sensitivity | Detection limit of immunohistochemistry is 10-3 | The detection limit of immunoblotting is 10-5 |
| Scope of application | Including other animal transmissible spongiform encephalopathies | Focus on bovine-specific diagnosis |
Analysis of core diagnostic technologies
1. H·E histopathological staining
The optimized operation process emphasizes:
- Formic acid treatment time is shortened to 1 hour to reduce tissue damage
- Paraffin immersion adopts gradient temperature control (56℃→60℃)
- Vacuolar lesions must meet the bilateral symmetric distribution characteristics
2. Immunohistochemical method
Key upgrade points include:
A provincial laboratory found that when using standard process testing:
① Proteinase K digestion time needs to be strictly controlled within 15±1 minutes
② High-pressure antigen retrieval requires the use of special ceramic containers to avoid buffer pH drift
3. New immunoblotting method
Technical points:
- The sampling weight of brain occlusal tissue is accurate to 100-130mg
- Quantitative control of PK enzyme final concentration ≥50μg/mL
- Positive judgment requires the simultaneous appearance of three characteristic bands of 17-33ku
Implementation suggestions
Laboratories should establish:
- Negative pressure operation area above biosafety level 2
- Traceability system for reference materials (such as BSE positive control provided by OIE)
- Multi-method joint inspection mechanism: Western blotting + histopathology double verification
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