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BS EN ISO 15216-1:2017+A1:2021

BS EN ISO 15216-1:2017+A1:2021

$198.66

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

Published By Publication Date Number of Pages
BSI 2021 54
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This document specifies a method for the quantification of levels of HAV and norovirus genogroup I (GI) and II (GII) RNA, from test samples of foodstuffs (soft fruit, leaf, stem and bulb vegetables, bottled water, BMS) or food surfaces. Following liberation of viruses from the test sample, viral RNA is then extracted by lysis with guanidine thiocyanate and adsorption on silica. Target sequences within the viral RNA are amplified and detected by 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, food surfaces or other matrices.

PDF Catalog

PDF Pages PDF Title
2 National foreword
4 European foreword
Anchor 2
Foreword to amendment A1
7 Foreword
8 Introduction
9 1 Scope
2 Normative references
3 Terms and definitions
11 4 Principle
4.1 Virus extraction
12 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
13 4.5 Test results
5 Reagents
5.1 General
5.2 Reagents used as supplied
14 5.3 Prepared reagents
15 6 Equipment and consumables
16 6.10 Microcentrifuge.
17 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
18 8.2.5 Bottled water
19 8.2.6 Bivalve molluscan shellfish
8.3 RNA extraction
20 8.4 Real-time RT-PCR
8.4.1 General requirements
8.4.2 Real-time RT-PCR analysis
22 9 Interpretation of results
9.1 General
9.2 Construction of standard curves
23 9.3 Calculation of RT-PCR inhibition
9.4 Calculation of extraction efficiency
24 9.5 Sample quantification
25 10 Expression of results
11 Precision
11.1 Interlaboratory study
11.2 Repeatability
11.3 Reproducibility limit
26 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)
34 Annex E (informative) Growth of mengo virus strain MC0 for use as a process control
35 Annex F (informative) RNA extraction using the NucliSENS® system
37 Annex G (informative) Generation of dsDNA control stocks
40 Annex H (informative) Generation of EC RNA stocks
42 Annex I (informative) Typical optical plate layout
44 Annex J (informative) Method validation studies and performance characteristics
53 Bibliography

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BS EN ISO 15216-1:2017+A1:2021
$198.66