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eLibrary > BS EN ISO 15216-1:2017 - TC Tracked Changes. Microbiology of the food chain. Horizontal method for determination of hepatitis A virus and norovirus using real-time RT-PCR - Method for quantification >

BS EN ISO 15216-1:2017 - TC Tracked Changes. Microbiology of the food chain. Horizontal method for determination of hepatitis A virus and norovirus using real-time RT-PCR - Method for quantification, 2020

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BS EN ISO 15216-1:2017 - TC Tracked Changes. Microbiology of the food chain. Horizontal method for determination of hepatitis A virus and norovirus using real-time RT-PCR - Method for quantification, 2020

30311174.pdf [Go to Page]
Foreword
Introduction
1 Scope
2 Normative references
3 Terms and definitions
4 Principle
4.1 Virus extraction
4.2 RNA extraction
4.3 Real-time RT-PCR
4.4 Control materials
4.4.1 Process control virus
4.4.2 Double-stranded DNA (dsDNA) control
4.4.3 EC RNA control
4.5 Test results
5 Reagents
5.1 General
5.2 Reagents used as supplied
5.3 Prepared reagents
6 Equipment and consumables
7 Sampling
8 Procedure
8.1 General laboratory requirements
8.2 Virus extraction
8.2.1 Process control virus material
8.2.2 Negative process control
8.2.3 Food surfaces
8.2.4 Soft fruit, leaf, stem and bulb vegetables
8.2.5 Bottled water
8.2.6 Bivalve molluscan shellfish
8.3 RNA extraction
8.4 Real-time RT-PCR
8.4.1 General requirements
8.4.2 Real-time RT-PCR analysis
9 Interpretation of results
9.1 General
9.2 Construction of standard curves
9.3 Calculation of RT-PCR inhibition
9.4 Calculation of extraction efficiency
9.5 Sample quantification
10 Expression of results
11 Precision
11.1 Interlaboratory study
11.2 Repeatability
11.3 Reproducibility limit
12 Test report
Annex A (normative) Diagram of procedure
Annex B (normative) Composition and preparation of reagents and buffers
Annex C (informative) Real-time RT-PCR mastermixes and cycling parameters
Annex D (informative) Real-time RT-PCR primers and hydrolysis probes for the detection of HAV, norovirus GI and GII and mengo virus (process control)
Annex E (informative) Growth of mengo virus strain MC0 for use as a process control
Annex F (informative) RNA extraction using the NucliSENS® system
Annex G (informative) Generation of dsDNA control stocks
Annex H (informative) Generation of EC RNA stocks
Annex I (informative) Typical optical plate layout
Annex J (informative) Method validation studies and performance characteristics
Bibliography [Go to Page]

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BS EN ISO 15216-1:2017 - TC Tracked Changes. Microbiology of the food chain. Horizontal method for determination of hepatitis A virus and norovirus using real-time RT-PCR - Method for quantification, 2020

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