GB/T 40253-2021 in English
VALIDCode of diagnosis for mikrocytosis of oysters—Microscopic methods
- Issued on:2021-05-21
- Implemented on:2021-12-01
- File Format:PDF
- Delivery:Via email within 1~3 business days
$214.00
《GB/T 40253-2021牡蛎小胞虫病诊断规程 显微镜检查组织法》由TC156（全国水产标准化技术委员会）归口，TC156SC11（全国水产标准化技术委员会水产养殖病害防治分会）执行，主管部门为农业农村部。
Introduction
Standard Technical Background
GB/T40253-2021 is my country's first national standard specifically for the microscopic diagnosis of oyster microcystis disease, filling the gap in the standard field of shellfish parasite detection. The standard is based on the requirements of the OIE Aquatic Animal Health Code and is formulated in combination with the characteristics of my country's aquaculture.
Comparison of core detection methods
| Technical indicators | Tissue printing method | Histopathological method |
|---|---|---|
| Detection sensitivity | 80-85% | 90-95% |
| Operation time | 2-3 hours | 24-48 hours |
| Observation of insect body characteristics | Basic morphological identification | Tissue localization + development stage determination |
Key technical points
1. Standardized operation of tissue printing method
Key control points: The thickness of the printed slide should maintain a single cell layer, and the Giemsa staining time should be strictly controlled within 30±2 minutes. The typical body characteristics are:
- Oval in diameter 2-4μm
- Cytoplasm blue/light blue (staining difference)
- Cell nucleus red
2. Optimization of the histopathological process
Attention should be paid to the dehydration and transparency step:
- The time error of each step of ethanol gradient dehydration is ≤5 minutes
- The tissue block should be translucent when xylene is transparent
- The paraffin infiltration temperature should be controlled at 56±1℃
Implementation suggestions
1. Sampling strategy
Give priority to adductor muscle and mantle tissues. Sampling in winter (water temperature <10℃) can increase the detection rate by more than 30%.
2. Cross-validation of results
It is recommended that positive samples be confirmed by transmission electron microscopy (SC/T7209.3-2007), especially for the first detection of cases in non-epidemic areas.
3. Laboratory quality control
For each batch of testing, the following should be set up:
- Positive control (known infected tissue)
- Negative control (healthy oyster tissue)
- Reagent blank (PBS replacement)
Technology evolution analysis
Compared with the 2007 version of the industry standard, this version has the following major improvements:
- Added the formula of DAFA fixative (Appendix A)
- Refine the ethanol gradient preparation specifications (A.4-A.8)
- Clarify the criteria for judging the worm development stage (Appendix C)
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