EN ISO 24914:2026

SLOVENSKI STANDARD
oSIST prEN ISO 24914:2025
01-junij-2025
Mikrobiologija v prehranski verigi - Z zanko posredovana izotermna amplifikacija
(LAMP) za odkrivanje mikroorganizmov in z njimi povezanih genetskih
označevalcev - Splošne zahteve in definicije (ISO/DIS 24914:2025)
Microbiology of the food chain - Loop-mediated isothermal amplification (LAMP) for the
detection of microorganisms and associated genetic markers - General requirements
and definitions (ISO/DIS 24914:2025)
Mikrobiologie der Lebensmittelkette - Loop-mediated isothermal amplification (LAMP)
zum Nachweis von Mikroorganismen - Allgemeine Anforderungen und Leitlinien
(ISO/DIS 24914:2025)
Microbiologie de la chaîne alimentaire - Amplification isotherme à médiation par boucle
(LAMP) pour la détection de micro-organismes et de marqueurs génétiques associés -
Exigences générales et définitions (ISO/DIS 24914:2025)
Ta slovenski standard je istoveten z: prEN ISO 24914
ICS:
07.100.30 Mikrobiologija živil Food microbiology
oSIST prEN ISO 24914:2025 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

oSIST prEN ISO 24914:2025
oSIST prEN ISO 24914:2025
DRAFT
International
Standard
ISO/DIS 24914
ISO/TC 34/SC 9
Microbiology of the food chain —
Secretariat: AFNOR
Loop-mediated isothermal
Voting begins on:
amplification (LAMP) for the
2025-04-14
detection of microorganisms
Voting terminates on:
and associated genetic markers
2025-07-07
— General requirements and
definitions
ICS: 07.100.30
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENTS AND APPROVAL. IT
IS THEREFORE SUBJECT TO CHANGE
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Reference number
ISO/DIS 24914:2025(en)
oSIST prEN ISO 24914:2025
DRAFT
ISO/DIS 24914:2025(en)
International
Standard
ISO/DIS 24914
ISO/TC 34/SC 9
Microbiology of the food chain —
Secretariat: AFNOR
Loop-mediated isothermal
Voting begins on:
amplification (LAMP) for the
detection of microorganisms
Voting terminates on:
and associated genetic markers
— General requirements and
definitions
ICS: 07.100.30
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENTS AND APPROVAL. IT
IS THEREFORE SUBJECT TO CHANGE
AND MAY NOT BE REFERRED TO AS AN
INTERNATIONAL STANDARD UNTIL
PUBLISHED AS SUCH.
This document is circulated as received from the committee secretariat.
IN ADDITION TO THEIR EVALUATION AS
BEING ACCEPTABLE FOR INDUSTRIAL,
© ISO 2025
TECHNOLOGICAL, COMMERCIAL AND
USER PURPOSES, DRAFT INTERNATIONAL
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
STANDARDS MAY ON OCCASION HAVE TO
ISO/CEN PARALLEL PROCESSING
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BE CONSIDERED IN THE LIGHT OF THEIR
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TO SUBMIT, WITH THEIR COMMENTS,
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Website: www.iso.org
Published in Switzerland Reference number
ISO/DIS 24914:2025(en)
ii
oSIST prEN ISO 24914:2025
ISO/DIS 24914:2025(en)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 2
4 Principle . 3
4.1 General .3
4.2 Laboratory procedure .4
4.3 Data interpretation .4
4.4 Performance criteria .4
5 General laboratory guidance . 4
5.1 General .4
5.2 Laboratory setup .5
5.3 Use of controls .5
5.3.1 General .5
5.3.2 Positive extraction control .6
5.3.3 Internal/external amplification control .6
6 Reagents and consumables . 6
7 Equipment . 7
7.1 General .7
7.1.1 Isothermal apparatus.7
7.1.2 Systems for detection of LAMP amplicons, comprising .7
8 Laboratory procedure . 7
8.1 General .7
8.2 Sample preparation .7
8.3 Nucleic acid extraction .7
8.4 Isothermal amplification .8
8.5 Signal detection .8
9 Data interpretation . 8
9.1 General .8
9.2 Interpretation of controls .8
9.3 Interpretation of test portion results .9
9.4 Test report .9
10 Performance characteristics . 9
Annex A (informative) LAMP basics . 10
Bibliography . 14

iii
oSIST prEN ISO 24914:2025
ISO/DIS 24914:2025(en)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards
bodies (ISO member bodies). The work of preparing International Standards is normally carried out through
ISO technical committees. Each member body interested in a subject for which a technical committee
has been established has the right to be represented on that committee. International organizations,
governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely
with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
The procedures used to develop this document and those intended for its further maintenance are 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 editorial rules of the
ISO/IEC Directives, Part 2 (see www.iso.org/directives).
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of any patent
rights identified during the development of the document will be in the Introduction and/or on the ISO list of
patent declarations received (see www.iso.org/patents).
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation of the voluntary nature of standards, the meaning of ISO specific terms and expressions
related to conformity assessment, as well as information about ISO's adherence to the World Trade
Organization (WTO) principles in the Technical Barriers to Trade (TBT), see www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 9,
Microbiology, in collaboration with the European Committee for Standardization (CEN) Technical Committee
CEN/TC 463, Microbiology of the food chain, in accordance with the Agreement on technical cooperation
between ISO and CEN (Vienna Agreement).
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 24914:2025
ISO/DIS 24914:2025(en)
Introduction
Loop-mediated isothermal amplification (LAMP) is used for the detection of microorganisms and associated
genetic markers (e.g. antimicrobial resistance genes, virulence genes) in the food chain.
This document provides the general requirements and guidance for the development and application of
LAMP-based methods for the detection of microorganisms and associated genetic markers sampled along
the food chain (e.g. ingredients, human food, animal food, and the production environment), and includes
sample preparation, nucleic acid extraction, isothermal amplification, signal detection, data interpretation,
report generation, and performance criteria. The document is applicable to all LAMP methods, platforms,
items from the food chain and associated laboratories. This document can be used by academia, industry,
and regulatory bodies for developing and applying LAMP methods for the detection of microorganisms and
associated genetic markers in the food chain.

v
oSIST prEN ISO 24914:2025
oSIST prEN ISO 24914:2025
DRAFT International Standard ISO/DIS 24914:2025(en)
Microbiology of the food chain — Loop-mediated isothermal
amplification (LAMP) for the detection of microorganisms
and associated genetic markers — General requirements and
definitions
WARNING — In order to protect the health and safety of laboratory personnel and the environment, it
is essential that tests for the detection of microorganisms are only undertaken in properly equipped
laboratories and under the control of skilled laboratory personnel. Appropriate precautions shall
be taken in the disposal of all samples and cultured materials. Persons using this document shall
be familiar with normal laboratory safety practices. This document does not purport to address all
of the health and safety aspects, if any, associated with its use. It is the responsibility of the user to
establish appropriate health and safety practices.
1 Scope
This document specifies the general requirements and provides guidance for the development and
application of loop-mediated isothermal amplification (LAMP) to detect microorganisms and associated
genetic markers (e.g. antimicrobial resistance genes, virulence genes) in the food chain. This document is
applicable to all LAMP methods, platforms, items from food chain and laboratories. The use of LAMP for
quantification is out of scope for this document.
Validation and verification of LAMP methods as either alternative or reference methods are not covered in
this document. Both validation and verification of microbiological methods are discussed in detail in the
ISO 16140 series and ISO 17468.
General requirements for isothermal methods including LAMP for molecular biomarker analysis are
covered by ISO 22942-1, and general requirements and definitions for polymerase chain reaction (PCR) for
the detection and quantification of microorganisms in the food chain are covered by ISO 22174.
This document has been established for microorganisms in the food chain and is applicable to:
— products intended for human consumption;
— products for feeding animals;
— environmental samples in the area of food and feed production and handling;
— samples from the primary production stage for the above items.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content constitutes
requirements of this document. For dated references, only the edition cited applies. For undated references,
the latest edition of the referenced document (including any amendments) applies.
ISO 6887 (all parts), Microbiology of the food chain — Preparation of test samples, initial suspension and
decimal dilutions for microbiological examination
ISO 7218, Microbiology of the food chain — General requirements and guidance for microbiological examinations
ISO 22174, Microbiology of the food chain — Polymerase chain reaction (PCR) for the detection and quantification
of microorganisms — General requirements and definitions

oSIST prEN ISO 24914:2025
ISO/DIS 24914:2025(en)
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at https:// www .electropedia .org/
3.1
loop-mediated isothermal amplification
LAMP
isothermal nucleic acid amplification technique that relies on auto-cycling strand-displacement
deoxyribonucleic acid (DNA) synthesis that is performed by a DNA polymerase with high strand-
displacement activity and a set of two specially designed inner and two outer primers to form a stem-loop
DNA structure during initiation steps, that serves as the starting material for second-stage LAMP cycling
where a set of two (sometimes one) loop primers may be added to speed up amplification
3.2
reverse transcription-loop-mediated isothermal amplification
RT-LAMP
method consisting of two reactions, a reverse transcription (RT) of ribonucleic acid (RNA) to single-stranded
complementary deoxyribonucleic acid (cDNA), followed by a loop-mediated isothermal amplification (LAMP) (3.1)
Note 1 to entry: One-step RT-LAMP is performed in a single tube. Two-step RT-LAMP can either be performed
sequentially in a single tube or in two different tubes.
[SOURCE: ISO 22174:2024, 3.3.3, modified — “LAMP” replaced “PCR” in the term name, domain, and Note 1
to entry.]
3.3
strand-displacing DNA polymerase
deoxyribonucleic acid (DNA) polymerase used for DNA amplification characterized by DNA template-
dependent 5’→3’ polymerase activity and strand displacement activity at a single elevated temperature (e.g.
between 60 °C and 65 °C), that also lacks 5’→3’ exonuclease activity
Note 1 to entry: strand-displacing DNA polymerase is commonly used for DNA amplification by LAMP.
3.4
positive extraction control
sample, spiked with a microorganism, treated in the same way as the test samples from extraction to
amplification to monitor the extraction process
[SOURCE: ISO 22174:2024, 3.5.2, modified — “from extraction to amplification to monitor the extraction
process” replaced “to monitor the entire process of the polymerase-chain-reaction-based method.”]
3.5
internal amplification control
nucleic acid added to each reaction in a defined amount or copy number which serves as an internal control
for amplification
Note 1 to entry: This nucleic acid sequence can be endogenous (naturally present in the tested matrix) or exogenous
(naturally absent in the tested matrix).
Note 2 to entry: An exogenous internal amplification control can be homologous (amplified using the same primers
as used for amplification of the target) or heterologous (amplified using different primers than those used for
amplification of the target). A homologous internal amplification control amplicon shall be distinguishable from the
microbial target amplicon (e.g. by melting temperature).
[SOURCE: ISO 22174:2024, 3.5.5, modified — “by melting temperature” replaced “by size or by insertion of a
different probe-binding sequence.”]

oSIST prEN ISO 24914:2025
ISO/DIS 24914:2025(en)
3.6
positive loop-mediated isothermal amplification control
positive LAMP control
loop-mediated isothermal amplification (LAMP) (3.1) reaction containing the target nucleic acid in a defined
amount or copy number
[SOURCE: ISO 22174:2024, 3.5.7, modified — “LAMP” replaced “PCR” in the term name, added “loop-mediated
isothermal amplification (LAMP) (3.1)” in the domain.]
3.7
negative loop-mediated isothermal amplification control
negative LAMP control
loop-mediated isothermal amplification (LAMP) (3.1)control made with water (or other LAMP-inert substrate
such as elution buffer) free of target nucleic acid and LAMP inhibitors
Note 1 to entry: Also referred to as no-template control (NTC).
[SOURCE: ISO 22174:2024, 3.5.8, modified — “LAMP” replaced “PCR” in the term name and domain, deleted
“grinding or.”]
3.8
loop-mediated isothermal amplification inhibitor
LAMP inhibitor
component of the test portion that negatively interferes with the nucleic acid amplification of loop-mediated
isothermal amplification (LAMP) (3.1) resulting in a null or partial amplification of the nucleic acid target, or
that reduces signal generation
Note 1 to entry: Examples of inhibitors include bile salts, calcium chloride, and humic acid.
[SOURCE: see References [9-12]]
3.9
non-laboratory field setting
workspace lacking conditions controlled for environmental aerosol contamination and sophisticated nucleic
acid purification apparatus
[SOURCE: ISO 16577:2022, 3.3.41]
4 Principle
4.1 General
There are four to six specially designed LAMP primers, that target six to eight regions of the template. The
amplification strategy of LAMP is based on the formation of a stem-loop DNA structure during initial steps
that serves as the starting material for second-stage LAMP cycling. Unlike PCR that relies on thermal cycling
to denature DNA and enable amplification by Taq DNA polymerase or equivalent, LAMP uses a strand-
displacing DNA polymerase (e.g. Bst DNA polymerase large fragment from the thermophilic bacterium
Geobacillus stearothermophilus [previously Bacillus stearothermophilus, hence Bst]), that allows auto-cycling
amplification under a constant temperature (e.g. between 60 °C and 65 °C). LAMP amplifies the target nucleic
acid sequence efficiently, with approximately 10 copies generated within an hour. The amplicon detection
chemistries for LAMP are versatile, and include colorimetry, fluorescence, bioluminescence, and turbidity,
among others. Platforms such as a colorimetric, turbidity, or fluorescence detection instrument, lateral
flow device or microfluidic apparatus may be employed. Descriptions of LAMP primer design, amplification
strategy, and detection chemistry can be found in Annex A.
LAMP-based methods are rapid (detection within 1 hour or as soon as 5 minutes), sensitive, specific, robust
(tolerance to inhibitors with simplified nucleic acid extraction), versatile (a number of detection chemistries/
platforms), and field-amenable (portable instrument). However, multiplex detection may be a challenge for
LAMP-based methods and there is a high risk of contamination if LAMP reaction tubes are opened post-
amplification in areas close to those used to perform LAMP.

oSIST prEN ISO 24914:2025
ISO/DIS 24914:2025(en)
The entire process can include the following steps:
a) sample preparation;
b) nucleic acid extraction;
c) isothermal amplification;
d) signal detection;
e) data interpretation.
4.2 Laboratory procedure
Sampling is out of scope for this document. Follow the specific International Standard dealing with sampling,
e.g. ISO 7218 gives guidance on sampling certain products. If there is no specific International Standard
dealing with the sampling of the product, the concerned parties should come to an agreement on this subject.
The laboratory procedure should start with sample preparation including the enrichment or concentration
of microorganisms and associated genetic markers from the test portion.
Any sample along the food chain is suitable as a test portion, provided it is possible to retrieve nucleic
acid (DNA and/or RNA) from the test portion if it is present and provided it has been established that the
nucleic acid solution prepared from the test portion does not significantly inhibit the LAMP assay (either
amplification or detection). Appropriate controls should be included (see Subclause 5.3).
Sample preparation is followed by nucleic acid extraction. Microbial cells in the test portion or enriched
culture are lysed to release their nucleic acid (DNA and/or RNA). If required, a separation stage is included
prior to lysis and/or a purification step is carried out following lysis.
LAMP amplification of the target nucleic acid sequence shall be performed using specific primers with the
addition of colorimetric substrates, fluorescent dyes, or other chemistries, under isothermal conditions in a
suitable instrument. The detection of LAMP amplicon is achieved through
...