FprEN ISO 17174
(Main)Molecular biomarker analysis - DNA barcoding of fish and fish products using defined mitochondrial cytochrome b and cytochrome c oxidase I gene segments (ISO/FDIS 17174:2024)
Molecular biomarker analysis - DNA barcoding of fish and fish products using defined mitochondrial cytochrome b and cytochrome c oxidase I gene segments (ISO/FDIS 17174:2024)
This document describes a procedure for the identification of single fish and fish fillets to the
level of genus or species.
The identification of fish species is carried out by PCR amplification of either a segment of the
mitochondrial cytochrome b gene (cytb) [1] or the cytochrome c oxidase I gene (cox1, syn COI)
[2], [3] or both, followed by sequencing of the PCR products and subsequent sequence
comparison with entries in databases [4], [5]. The methodology allows the identification of a large
number of commercially important fish species.
The decision whether the cytb or cox1 gene segment or both are used for fish identification
depends on the declared fish species, the applicability of the PCR method for the fish species
and the availability of comparative sequences in the public databases.
This method has been successfully validated on raw fish fillets, however, laboratory experience
is available that it can also be applied to processed, e.g. cold smoked, hot smoked, salted,
frozen, cooked, fried, deep fried
samples.
This document is usually unsuitable for the analysis of highly processed foods, e.g. tins of fish,
with highly degraded DNA where the fragment lengths are not sufficient for amplification of the
targets. Furthermore, it is not applicable for complex fish products containing mixtures of two or
more fish species.
Untersuchung auf molekulare Biomarker - DNA-Barcoding von Fisch und Fischprodukten anhand definierter mitochondrialer Cytochrom b- und Cytochrom c-Oxidase I-Genabschnitte (ISO/FDIS 17174:2024)
Dieses Dokument legt ein Verfahren für die Identifizierung von einzelnen Fischen und Fischfilets bis zur Ebene der Gattung oder Art fest. Es ermöglicht die Identifizierung einer großen Anzahl kommerziell wichtiger Fischarten mittels DNA-Barcoding.
Dieses Verfahren wurde an rohem Fisch validiert. Laborerfahrungen deuten auf eine zusätzliche Anwendbarkeit auf verarbeitete Fischerzeugnisse hin (z. B. kalt geräucherte, heiß geräucherte, gesalzene, tiefgefrorene, gekochte, gebratene und frittierte Proben).
Das beschriebene Verfahren ist in der Regel ungeeignet für die Analyse stark verarbeiteter Lebensmittel (z. B. Fischkonserven, mit stark degradierter DNA, bei denen die Fragmentlängen nicht für eine Amplifikation der Zielsequenzen (Targets) ausreichen). Außerdem gilt es nicht für zusammengesetzte Fischerzeugnisse, die Mischungen aus zwei oder mehr Fischarten enthalten.
Die Identifizierung der Fischarten erfolgt durch PCR-Amplifikation, entweder eines Segments des mitochondrialen Cytochrom-b-Gens (cytb) oder des Cytochrom-c-Oxidase-I-Gens (cox1, syn COI) oder von beiden, gefolgt von der Sequenzierung der PCR-Produkte und einem anschließenden Sequenzvergleich mit Einträgen in Datenbanken.
Analyse de biomarqueurs moléculaires - Codes-barres d’ADN de poissons et de produits à base de poisson à l’aide de segments de gènes mitochondriaux de cytochrome b et cytochrome c oxydase I (ISO/FDIS 17174:2024)
Analiza molekularnih biomarkerjev - Črtno kodiranje DNK rib in ribjih proizvodov z uporabo definiranih mitohondrijskih segmentov genov citokrom b in citokrom c oksidaze I (ISO/FDIS 17174:2024)
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
oSIST prEN ISO 17174:2023
01-julij-2023
Analiza molekularnih biomarkerjev - Črtno kodiranje DNK rib in ribjih proizvodov z
uporabo segmentov genov, ki nosijo zapis za mitohondrijski citokrom b in
citokrom c oksidaze I (ISO/DIS 17174:2023)
Molecular biomarker analysis - DNA barcoding of fish and fish products using defined
mitochondrial cytochrome b and cytochrome c oxidase I gene segments (ISO/DIS
17174:2023)
Untersuchung auf molekulare Biomarker - DNA-Barcoding von Fisch und Fischprodukten
anhand definierter mitochondrialer Cytochrom b- und Cytochrom c-Oxidase I-
Genabschnitte (ISO/DIS 17174:2023)
Analyse de biomarqueurs moléculaires - Codes-barres d’ADN de poissons et de produits
à base de poisson à l’aide de segments de gènes mitochondriaux de cytochrome b et
cytochrome c oxydase I (ISO/DIS 17174:2023)
Ta slovenski standard je istoveten z: prEN ISO 17174
ICS:
67.120.30 Ribe in ribji proizvodi Fish and fishery products
oSIST prEN ISO 17174:2023 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
oSIST prEN ISO 17174:2023
oSIST prEN ISO 17174:2023
DRAFT INTERNATIONAL STANDARD
ISO/DIS 17174
ISO/TC 34/SC 16 Secretariat: ANSI
Voting begins on: Voting terminates on:
2023-05-15 2023-08-07
Molecular biomarker analysis — DNA barcoding of fish and
fish products using defined mitochondrial cytochrome b
and cytochrome c oxidase I gene segments
ICS: 67.120.30
This document is circulated as received from the committee secretariat.
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NATIONAL REGULATIONS.
ISO/DIS 17174:2023(E)
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PROVIDE SUPPORTING DOCUMENTATION. © ISO 2023
oSIST prEN ISO 17174:2023
DRAFT INTERNATIONAL STANDARD
ISO/DIS 17174
ISO/TC 34/SC 16 Secretariat: ANSI
Voting begins on: Voting terminates on:
Molecular biomarker analysis — DNA barcoding of fish and
fish products using defined mitochondrial cytochrome b
and cytochrome c oxidase I gene segments
ICS: 67.120.30
This document is circulated as received from the committee secretariat.
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENT AND APPROVAL. IT IS
© ISO 2023
ISO/CEN PARALLEL PROCESSING
THEREFORE SUBJECT TO CHANGE AND MAY
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
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ii
PROVIDE SUPPORTING DOCUMENTATION. © ISO 2023
oSIST prEN ISO 17174:2023
ISO/DIS 17174:2023(E)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 4
5 Reagents and materials . 4
5.1 General . 4
5.2 PCR reagents . 4
6 Apparatus . 5
7 Procedure .5
7.1 Sample preparation . 5
7.2 DNA extraction . 6
7.3 PCR . 6
7.3.1 General . 6
7.3.2 PCR setup . 6
7.3.3 Temperature-time program . 7
7.3.4 PCR controls . 8
8 Evaluation . 8
8.1 Evaluation of PCR products . . 8
8.2 Evaluation of the PCR results . 8
8.3 Sequencing of PCR products . 9
8.4 Evaluation of sequence data . 9
8.5 Comparison of the sequence with public databases . 9
8.5.1 General . 9
8.5.2 Sequence comparison of cytb and/or cox1 DNA sequences with GenBank . 10
8.5.3 Sequence comparison of cox1 DNA sequences with BOLD. 10
9 Interpretation of database query results .11
10 Validation status and performance criteria .12
10.1 Collaborative study for the identification of fish species based on cytb sequence
analysis .12
10.2 Collaborative study for the identification of fish species based on cox1 sequence
analysis .13
11 Test report .14
Annex A (informative) Practical laboratory experiences with the amplifiability of cytb and
cox1 segments from tested fish species.15
Bibliography .18
iii
oSIST prEN ISO 17174:2023
ISO/DIS 17174:2023(E)
Foreword
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electrotechnical standardization.
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described in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the
different types of ISO documents should be noted. This document was drafted in accordance with the
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Attention is drawn to the possibility that some of the elements of this document may be the subject of
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www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee 34. Food Products, Subcommittee SC 16,
Horizontal methods for molecular biomarker analysis.
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html.
iv
oSIST prEN ISO 17174:2023
ISO/DIS 17174:2023(E)
Introduction
Food safety is a key aspect in terms of consumer protection. In the last three decades, globalization has
taken place in the trade of food. Fish trade channels are becoming steadily longer and more complicated
so that sophisticated traceability tools are needed to ensure food safety. Correct food labelling is
a prerequisite to ensure safe fish products and fair trade as well as to minimize illegal, unreported
and unregulated (IUU) fishing. In particular, the fact that fish is increasingly being processed in
export countries makes the identification of species by morphological characteristics impossible.
The development of harmonized and standardized protocols for the authentication of fish products is
necessary to establish reliable methods for the detection of potential food fraud.
v
oSIST prEN ISO 17174:2023
oSIST prEN ISO 17174:2023
DRAFT INTERNATIONAL STANDARD ISO/DIS 17174:2023(E)
Molecular biomarker analysis
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