GB/T 18935-2018 in English
VALIDDiagnostic techniques for foot and mouth disease
- Issued on:2018-09-17
- Implemented on:2019-04-01
- File Format:PDF
- Delivery:Via email within 5 business days
$418.00
《GB/T 18935-2018口蹄疫诊断技术》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/T18935-2018 is the core technical standard for the diagnosis of foot-and-mouth disease in my country. Compared with the 2003 version, it has undergone the following technical upgrades:
| Technical Module | Content of the 2003 version | Updates to the 2018 version |
|---|---|---|
| Clinical Diagnosis | Not an independent chapter | Newly added susceptible animal determination and symptom grading system |
| Serological Testing | VIA-AGID | Replaced with 3ABC-I/B-ELISA |
| Molecular diagnosis | Basic RT-PCR | Newly added stereotyped RT-PCR and fluorescent quantitative RT-PCR |
Analysis of core diagnostic technologies
1. Key points of virus isolation technology
The standard stipulates the use of a dual-track system of suckling mouse inoculation and cell culture:
- Suckling mouse inoculation: 0.2mL of the treated sample is subcutaneously inoculated on the back of the neck of 3-day-old suckling mice, and paralysis symptoms are observed within 72 hours
- Cell culture: IB-RS-2 (porcine source) or BHK-21 (universal) cell lines are preferred, and the CPE judgment threshold is set to ≥50%
2. Comparison of serological detection methods
| Method | Detection target | Sensitivity | Judgment standard |
|---|---|---|---|
| Typing ELISA | Viral antigen | 1:8 dilution | Relative OD value ≥0.3 |
| LPB-ELISA | Structural protein antibody | 1:128 | 50% endpoint titer |
| 3ABC-ELISA | Non-structural protein antibody | 0.3 titer | Distinguishing infection from immunity |
Application of molecular diagnostic technology
Fluorescence quantitative RT-PCR operation specifications
TaqMan probe method is used, and the recommended primer combination is:
Forward: CACYTYAAGRTGACAYTGRTACTGGTAC Reverse: CAGATYCCRAGT-GWCICITGTTA Probe: CCTCGGGGTACCTGAAGGGCATCC
Critical value: Ct value <35 is positive, and nuclease-free consumables must be used to prevent contamination
Recommendations for the implementation of the standard
- Laboratory hierarchical testing process: Clinical samples should first be screened by multiple RT-PCR, and positive samples should be typed
- Antibody testing should be combined with vaccination history, and positive 3ABC antibodies indicate wild virus infection
- VP1 gene sequencing is recommended to be sent to the OIE reference laboratory for comparison
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