Microbiology of food and animal feeding stuffs. Polymerase chain reaction (PCR) for the detection of food-borne pathogens. Requirements for amplification and detection for qualitative methods (ISO 20838:2006)
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ISO 20838:2006 provides the overall framework for qualitative methods for the detection of food-borne pathogens using the polymerase chain reaction (PCR). It covers the general requirements for the specific amplification of target nucleic acid sequences and the detection and confirmation of the identity of the amplified nucleic acid sequence. Guidelines, minimum requirements and performance characteristics described in ISO 20838:2006 are intended to ensure that comparable and reproducible results are obtained in different laboratories. ISO 20838:2006 has been established for food-borne pathogens in or isolated from food and feed matrices, but can also be applied to other matrices, for example environmental samples, or to the detection of other microorganisms under investigation.
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Sisällysluettelo
Foreword
Introduction
1 Scope
2 Normative references
3 Terms and definitions
4 Principle
5 Reagents
5.1 DNA polymerase
5.2 Reverse transcriptase
5.3 Reaction buffer
5.4 Deoxyribonucleoside triphosphates (dNTP), for PCR
5.5 Primers
5.6 Water
5.7 Magnesium chloride (MgCl2)
5.8 Chemicals for detection of PCR products
5.9 Optional additional reagents
6 Apparatus and equipment
7 Procedure
7.1 PCR amplification
7.2 Detection and/or confirmation of PCR products
7.3 Controls
7.4 General considerations for amplification reactions
ISO 16140:2003 Microbiology of food and animal feeding stuffs -- Protocol for the validation of alternative methods
ISO 22174:2005 Microbiology of food and animal feeding stuffs -- Polymerase chain reaction (PCR) for the detection of food-borne pathogens -- General requirements and definitions
ISO 16140:2003 Microbiology of food and animal feeding stuffs -- Protocol for the validation of alternative methods
ISO 22174:2005 Microbiology of food and animal feeding stuffs -- Polymerase chain reaction (PCR) for the detection of food-borne pathogens -- General requirements and definitions