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BS EN ISO 15216-2:2019

BS EN ISO 15216-2:2019

$198.66

Microbiology of the food chain. Horizontal method for determination of hepatitis A virus and norovirus using real-time RT-PCR – Method for detection

Published By Publication Date Number of Pages
BSI 2019 52
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This document specifies a method for detection of hepatitis A virus (HAV) and norovirus genogroups I (GI) and II (GII), from test samples of foodstuffs [(soft fruit, leaf, stem and bulb vegetables, bottled water, bivalve molluscan shellfish (BMS)] or surfaces using real-time RT-PCR.

This method is not validated for detection of the target viruses in other foodstuffs (including multi-component foodstuffs), or any other matrices, nor for the detection of other viruses in foodstuffs, surfaces or other matrices.

PDF Catalog

PDF Pages PDF Title
2 undefined
9 Foreword
10 Introduction
11 1 Scope
2 Normative references
3 Terms and definitions
13 4 Principle
4.1 Virus extraction
4.2 RNA extraction
4.3 Real-time RT-PCR
14 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
16 5.3 Reagents requiring preparation
17 6 Equipment and consumables
18 7 Sampling
8 Procedure
8.1 General laboratory requirements
8.2 Virus extraction
8.2.1 General
19 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
20 8.2.6 Bottled water
8.2.7 Bivalve molluscan shellfish (BMS)
21 8.3 RNA extraction
8.4 Real-time RT-PCR
8.4.1 General requirements
22 8.4.2 Real-time RT-PCR analysis
24 9 Interpretation of results
9.1 General
9.2 Construction of process control virus RNA standard curve
9.3 Control for RT-PCR inhibition
25 9.4 Calculation of extraction efficiency
10 Expression of results
26 11 Performance characteristics of the method
11.1 Validation study
11.2 Sensitivity
11.3 Specificity
11.4 LOD50
12 Test report
27 Annex A (normative) Diagram of procedure
28 Annex B (normative) Composition and preparation of reagents and buffers
31 Annex C (informative) Real-time RT-PCR mastermixes and cycling parameters
32 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)
35 Annex E (informative) Growth of mengo virus strain MC0 for use as a process control
36 Annex F (informative) RNA extraction using the BioMerieux NucliSens® system
38 Annex G (informative) Generation of external control RNA (EC RNA) stocks
41 Annex H (informative) Typical optical plate layout
42 Annex I (informative) Method validation studies and performance characteristics
50 Bibliography

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BS EN ISO 15216-2:2019
$198.66