GB/T 28630.3-2012Diagnostic protocols for white spot disease - Part 3: In situ hybridization method (English PDF)
白斑综合征(WSD)诊断规程 第3部分：原位杂交检测法
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Issued by
SAMR; SAC
Level / Type
National · Recommended
Issue date
July 31, 2012
Implementation date
November 1, 2012
Scope
GB/T 28630.3-2012 is the English-translated version of 白斑综合征(WSD)诊断规程 第3部分：原位杂交检测法.
GB/T 28630.3-2012 is the Chinese standard "Diagnostic protocols for white spot disease - Part 3: In situ hybridization method". Its scope clause reads: This section GB/T 28630 specifies the desired spot syndrome in situ hybridization assay reagents and materials, equipment, procedures, and Result judgment.
Assessment This section applies to conduct spot syndrome of shrimp and other sensitive host tissue cells and virus amplification degree of infection status And diagnosis of the disease.
Not suitable for carrying the virus non-infectious specimens for virus detection. Its clauses include reagents and materials.
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Document preview — GB/T 28630.3-2012
National Standard of the People's Republic of China
- ICS
- 65.020.30
- Classification
- B 41
Issued by: State Administration for Market Regulation; Standardization Administration of the PRC
Contents
- 1 Scope
- 2 Normative references
- 3.2 Technical Principle
- 4 Reagents and materials
- 4.7 neutral gum
1 Scope
This section GB/T 28630 specifies the desired spot syndrome in situ hybridization assay reagents and materials, equipment, procedures, and
Result judgment.
Assessment This section applies to conduct spot syndrome of shrimp and other sensitive host tissue cells and virus amplification degree of infection status
And diagnosis of the disease. Not suitable for carrying the virus non-infectious specimens for virus detection.
2 Normative references
The following documents for the application of this document is essential. For dated references, only the dated version suitable for use herein
Member. Who does not note date reference documents, the latest edition (including any amendments) applies to this document.
Diagnostic protocols GB/T 28630.1-2012 spot syndrome (WSD) - Part 1. nucleic acid probe dot blot hybridization assay
Diagnostic protocols GB/T 28630.4-2012 spot syndrome (WSD) Part 4. Histopathological diagnosis method
Principle 3
Pathogens and pathological features of White Spot Syndrome 3.1
See GB/T 28630.4-2012 2.1.
3.2 Technical Principle
In situ hybridization method is based on a non-radioactive marker --- digoxigenin (DIG) labeled white spot syndrome virus (WSSV) nucleic acid probe
Needle and slice WSSVDNA present in nucleic acid hybridization, and by an antigen - antibody reaction and enzyme - substrate chemical color reaction
It shows the presence of WSSV sites on tissue sections. In situ hybridization can be kept intact tissue and cell morphology in a complex organization can single
A cell for virus detection, accurately reflect the tissue distribution of the virus in cells.
4 Reagents and materials
4.1
Unless otherwise stated in the analysis using only recognized as analytical grade reagents and distilled or deionized water or equivalent purity.
4.2
nucleic acid probe according to GB/T 28630.1-2012 6.2 requirements.
4.3
DIG alkaline phosphatase conjugated antibody (AP- anti-DIG, 0.75U/µL, 4 °C). chemical reagents.
4.4
xylene.
4.5
ethanol.
4.7 neutral gum
Biological staining.
4.8
paraffin wax (melting point 52 °C ~ 54 °C). Slice.
4.9-Davidson's AFA (Davidson'sAFA) fixative. See Appendix A of the provisions.
......
This preview omits tables, figures, formulas and parts of the technical clauses. The complete document — 15 pages — is available in the English PDF.
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Related Standards
GB/T 28630.4-2012 — Diagnostic protocols for white spot disease - Part 4: Histopathology method
GB/T 28630.2-2012 — Diagnostic protocols for white spot disease - Part 2: Nested PCR method
GB/T 28630.5-2012 — Diagnostic protocols for white spot disease - Part 5: T-E staining method with fresh tissue
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GB/T 28630.3-2012
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