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BS EN ISO 15216-2:2019 Microbiology of the food chain. Horizontal method for determination of hepatitis A virus and norovirus using real-time RT-PCR - Method for detection >
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BS EN ISO 15216-2:2019 Microbiology of the food chain. Horizontal method for determination of hepatitis A virus and norovirus using real-time RT-PCR - Method for detection, 2019
- undefined
- 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 EC RNA control
- 4.5 Test results
- 5 Reagents
- 5.1 General
- 5.2 Reagents used as supplied
- 5.3 Reagents requiring preparation
- 6 Equipment and consumables
- 7 Sampling
- 8 Procedure
- 8.1 General laboratory requirements
- 8.2 Virus extraction
- 8.2.1 General
- 8.2.2 Process control virus material
- 8.2.3 Negative process control
- 8.2.4 Surfaces
- 8.2.5 Soft fruit and leaf, stem and bulb vegetables
- 8.2.6 Bottled water
- 8.2.7 Bivalve molluscan shellfish (BMS)
- 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 process control virus RNA standard curve
- 9.3 Control for RT-PCR inhibition
- 9.4 Calculation of extraction efficiency
- 10 Expression of results
- 11 Performance characteristics of the method
- 11.1 Validation study
- 11.2 Sensitivity
- 11.3 Specificity
- 11.4 LOD50
- 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 BioMerieux NucliSens® system
- Annex G (informative) Generation of external control RNA (EC RNA) stocks
- Annex H (informative) Typical optical plate layout
- Annex I (informative) Method validation studies and performance characteristics
- Bibliography [Go to Page]