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