GB/T 18645-2020 in English
VALIDDiagnostic techniques for animal tuberculosis
- Issued on:2020-12-14
- Implemented on:2020-12-14
- File Format:PDF
- Delivery:Via email within 5 business days
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《GB/T 18645-2020动物结核病诊断技术》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/T 18645-2020 replaces the 2002 version of the standard. The main technical changes include: new clinical diagnostic methods (epidemic characteristics, clinical symptoms, pathological changes), introduction of molecular biological detection (PCR technology), optimization of biosafety requirements (Appendix C regulates sampling and transportation), etc. The standard revision responds to OIE requirements and solves problems such as the long animal inoculation experiment cycle and inconsistent biochemical methods in the original standard.
Comparison of diagnostic method frameworks
| Method type | 2002 edition | 2020 edition | Improvement points |
|---|---|---|---|
| Bacteriological examination | Staining microscopy + isolation and culture | Add PNB/TCH biochemical identification | Standardized niacin test alternative |
| Intracutaneous allergy | Bovine PPD single test | Add avian PPD comparative test | Improve specificity |
| Molecular detection | Not included | Newly added class=instrument>PCR detection (bovine/tuberculosis/avian mycobacterium) | Detection limit of 103CFU/mL |
| Immunological detection | Not included | New in vitro detection of interferon-γ | Sensitivity of 92.3% |
Details of core detection technology
1. PCR detection process
Key steps: Sample pretreatment → DNA extraction (Extraction solution in Appendix D) → Amplification (primers in Table 2) → Electrophoresis analysis (characteristic bands such as 406bp/263bp/577bp). The detection limit of the IS901 gene is 15 copies/reaction.
2. Improvement of intradermal allergy
The double PPD comparison method (bovine PPD 2,000IU vs avian PPD 2,500IU) is used. The judgment criteria are:
- Positive: Δ skin thickness ≥ 4mm and bovine PPD reaction > avian PPD
- Suspicious: 2mm≤Δ skin thickness < 4mm
Implementation suggestions
Laboratory configuration: Class II biosafety cabinet, PCR instrument (ultraviolet disinfection procedure required), -80℃ refrigerator (nucleic acid storage).
Quality control: Each batch of PPD needs to be positive and negative controls, and PCR amplification needs to have an internal reference (such as β-actin).
Biosafety: Autopsy sampling must comply with the requirements of GB 19489, and Mycobacterium tuberculosis operations should be performed in a BSL-3 laboratory.
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