GB/T 35806-2018Animal influenza detection—Protocol of duplex real-time RT-PCR for influenza virus subtypes H7 and N9 (English PDF)
动物流感检测 H7N9亚型流感病毒双重荧光RT-PCR检测方法
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Issued by
SAMR; SAC
Level / Type
National · Recommended
Issue date
February 6, 2018
Implementation date
September 1, 2018
Scope
GB/T 35806-2018 is the English-translated version of 动物流感检测 H7N9亚型流感病毒双重荧光RT-PCR检测方法.
GB/T 35806-2018 is the Chinese national protocol for detecting the H7 and N9 subtypes of animal influenza virus in a single duplex real-time RT-PCR reaction. It sets out a TaqMan dual-fluorescence procedure that looks for H7 and N9 nucleic acid at the same time, and is meant for the rapid detection and screening of these subtypes in animals.
The document lists the abbreviations used, from Ct value, DEPC and the FAM, HEX and TAMRA dyes to the MGB probe, hemagglutinin and neuraminidase, and it specifies the reagents and materials, including TRIzol, pre-cooled chloroform and isopropyl alcohol and DEPC water prepared as given in Annex A. It relies on GB/T 19438.1-2004 and on GB 19489 for laboratory biosafety.
Veterinary and inspection laboratories use it. It was issued on 6 February 2018 and has applied since 1 September 2018.
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Document preview — GB/T 35806-2018
National Standard of the People's Republic of China
- ICS
- 11.220
- Classification
- B 41
Issued by: State Administration for Market Regulation; Standardization Administration of the PRC
Contents
- Foreword
- 1 Scope
- 2 Normative references
- 3 Abbreviations
- 4 Reagents and materials
Foreword
This standard was drafted in accordance with the rules given in GB/T 1.1-2009.
1 Scope
This standard specifies TaqMan dual-fluorescence RT-PCR procedures for the simultaneous detection of influenza A nucleic acid in H7 and N9 subtype animals. method.
This standard applies to the rapid detection and screening of animal influenza virus H7 and N9 subtypes.
2 Normative references
The following documents are indispensable for the application of this document. For dated references, only dated versions apply to this document. For undated references, the latest version (including all amendments) applies to this document.
GB/T 6682 Analysis Laboratory Water Specifications and Test Methods
GB/T 19438.1-2004 Avian influenza virus universal fluorescence RT-PCR detection method
GB 19489 General Requirements for Laboratory Biosafety
3 Abbreviations
The following abbreviations apply to this document.
Ct (or Cp) value. the number of cycles experienced by the amount of fluorescence signal in each reaction tube reaching the set threshold (ThresholdCycle, or CrossingPoint)
DEPC. Diethypyrocarbonate
FAM. Carboxyfluorescein
HA. Hemagglutinin
HEX. Hexachlorofluorescein
MGB. Minor GrooveBinder
NA. Neuraminidase
TAMRA. Carboxytetramethylrhodamine
Fluorescence RT-PCR. Real-time RT-PCR
4 Reagents and materials
4.1 Reagents
4.1.1 General. Unless otherwise stated, all reagents used are of analytical grade. All liquid reagents used shall be packed in RNase-free containers.
4.1.2 TRIzol. Store at 2°C to 25°C.
4.1.3 Chloroform. Precooling at 2°C to 8°C.
4.1.4 Isopropyl alcohol. Precooled at -20°C.
4.1.5 DEPC water. See A.1 for recipes.
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This preview omits tables, figures, formulas and parts of the technical clauses. The complete document — 15 pages — is available in the English PDF.
Referenced standards
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Related Standards
GB 19489-2008 — Laboratories - General requirements for biosafety
GB/T 19438.1-2004 — Method of the real-time RT-PCR for the detection of Avian Influenza Virus
GB/T 6682-2008 — Water for analytical laboratory use - Specification and test methods
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GB/T 35806-2018
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