ISO 21474-3:2024
(Main)In vitro diagnostic medical devices — Multiplex molecular testing for nucleic acids — Part 3: Interpretation and reports
In vitro diagnostic medical devices — Multiplex molecular testing for nucleic acids — Part 3: Interpretation and reports
This document gives the general requirements for interpretation and reporting of multiplex molecular tests which simultaneously identify two or more nucleic acid target sequences of interest. This document is applicable to all multiplex methods used for examination using in vitro diagnostic (IVD) medical devices and laboratory developed tests (LDTs). It provides information for both qualitative and quantitative detection of nucleic acid target sequences. This document is intended as guidance for multiplex examinations that detect or quantify human nucleic acid target sequences and microbial pathogen nucleic acid target sequences from human clinical specimens. This document is applicable to any molecular IVD examination performed by medical laboratories. It is also intended to be used by laboratory customers, IVD developers and manufacturers, biobanks, institutions, commercial organizations performing biomedical research, and regulatory authorities. This document is not applicable to metagenomic massive parallel sequencing (MPS), but it is applicable to multiplex molecular methods including 16S sequencing.
Dispositifs médicaux de diagnostic in vitro — Tests moléculaires multiplex pour les acides nucléiques — Partie 3: Interprétation et rapports
General Information
Standards Content (Sample)
International
Standard
ISO 21474-3
First edition
In vitro diagnostic medical
2024-11
devices — Multiplex molecular
testing for nucleic acids —
Part 3:
Interpretation and reports
Dispositifs médicaux de diagnostic in vitro — Tests moléculaires
multiplex pour les acides nucléiques —
Partie 3: Interprétation et rapports
Reference number
© ISO 2024
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ii
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 General requirements . 2
5 Interpretation of results . 2
5.1 General .2
5.2 Methods for interpretation of results .2
5.3 Documentation on bioinformatics analysis .3
5.4 Monitoring of bioinformatics analysis .4
5.5 Genomic databases .4
5.6 Reference sequence databases .5
5.7 Variant identification and annotation .5
5.8 Categorization of variants .6
6 Reporting of test results . 7
6.1 General .7
6.2 Reporting elements .7
6.3 Test report content .8
6.4 Reporting detected variants .9
6.5 Reporting of secondary findings .9
6.6 Reporting method .10
Annex A (informative) Multiplex molecular test for cancer .11
Annex B (informative) Multivariable molecular tests .12
Annex C (informative) Chromosome microarray analysis test reports .13
Bibliography . 14
iii
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards
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This document was prepared by Technical Committee ISO/TC 212, Medical laboratories and in vitro diagnostic
systems.
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Any feedback or questions on this document should be directed to the user’s national standards body. A
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iv
Introduction
The first generation of in vitro diagnostic (IVD) medical devices for nucleic acid-based molecular tests
has been focused on detection or quantitation of a single nucleic acid sequence (e.g. viral RNA, mRNA,
or genomic DNA) within a clinical specimen. By comparison, a multiplex molecular test simultaneously
measures multiple nucleic acid sequences of interest in a single reaction tube or a system. The development
and clinical use of multiplex IVD medical devices are rapidly expanding with the technological advances and
new elucidation of the clinical significance of many biomarkers.
In comparison to single target analysis, multiplex molecular tests require an increased number of controls,
more complex performance evaluation/data analysis algorithms, and more complex interpretation and
[1,2]
reporting of results. Some multiplex systems amplify multiple targets in a single reaction step and then
[3]
split these into reactions for specific target detection.
Laboratories can develop assays in-house (“laboratory-developed test (LDT)”, “home-brew”, or “in-house
test”) or use commercially available multiplex assays involving a variety of technologies and instrument
platforms. Multiplex molecular testing provides large amounts of complicated and multifarious genetic
information, resulting in significant challenges to the laboratory with regards to appropriate data analysis,
interpretation and reporting.
Implementation of a multiplex molecular test identifies large numbers of genetic variations in a sample,
which is crucial for optimal patient care, and treatment guidelines are developed based on specific molecular
findings; therefore, it is imperative to standardize the interpretation and reporting of molecular results
among laboratories performing these tests.
This document describes the requirements and recommendations for various aspects of interpretation and
reporting of the results by multiplex molecular tests in order to ensure the quality of laboratory services of
such tests, in implementing multiplex molecular nucleic acid tests for clinical use.
v
International Standard ISO 21474-3:2024(en)
In vitro diagnostic medical devices — Multiplex molecular
testing for nucleic acids —
Part 3:
Interpretation and reports
1 Scope
This document gives the general requirements for interpretation and reporting of multiplex molecular
tests which simultaneously identify two or more nucleic acid target sequences of interest. This document is
applicable to all multiplex methods used for examination using in vitro diagnostic (IVD) medical devices and
laboratory developed tests (LDTs). It provides information for both qualitative and quantitative detection of
nucleic acid target sequences.
This document is intended as guidance for multiplex examinations that detect or quantify human nucleic
acid target sequences and microbial pathogen nucleic acid target sequences from human clinical specimens.
This document is applicable to any molecular IVD examination performed by medical laboratories. It is also
intended to be used by laboratory customers, IVD developers and manufacturers, biobanks, institutions,
commercial organizations performing biomedical research, and regulatory authorities. This document is
not applicable to metagenomic massive parallel sequencing (MPS), but it is applicable to multiplex molecular
methods including 16S sequencing.
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 15189, Medical laboratories — Requirements for quality and competence
ISO 21474-1, In vitro diagnostic medical devices — Multiplex molecular testing for nucleic acids — Part 1:
Terminology and general requirements for nucleic acid quality evaluation
3 Terms and definitions
For the purposes of this document, terms and definitions given in ISO 21474-1 and the following 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
process step
part of a process which is predominantly self-sufficient and consists of one or several unit operations
[SOURCE: ISO 10209:2022, 3.1.65]
4 General requirements
Multiplex molecular tests are IVD and medical devices that measure multiple nucleic acid sequences
simultaneously, such as multiplex PCR, DNA microarray, and MPS-based methodologies.
A multivariable molecular test is a molecular test that combines the values of multiple variables using an
interpretation function to yield a single patient-specific result including “classification”, “score” and/or
[5]
“index”. This is usually based on a platform of multiplex molecular tests, e.g. a miRNA assay. For more
guidance, see Annex B.
An increasing number of clinical and commercial laboratories have been performing multiplex molecular
tests and issuing corresponding clinical reports to provide information for the care of their patients.
However, the detected variants and relevant information in each report can differ because of the use of
different methodologies (e.
...
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