SIST EN ISO 4307:2022

SLOVENSKI STANDARD
SIST EN ISO 4307:2022
01-januar-2022
Nadomešča:
SIST-TS CEN/TS 17305:2019
Molekularne diagnostične preiskave in vitro - Specifikacije za predpreiskovalne
procese za slino - Izolirana človeška DNK (ISO 4307:2021)
Molecular in vitro diagnostic examinations - Specifications for pre-examination processes
for saliva - Isolated human DNA (ISO 4307:2021)
Molekularanalytische in-vitro-diagnostische Verfahren - Spezifikationen für
präanalytische Prozesse für Speichel - Isolierte DNA (ISO 4307:2021)
Analyse de diagnostic moléculaire in vitro - Spécifications relatives aux processus
préanalytiques pour la salive - ADN humain extrait (ISO 4307:2021)
Ta slovenski standard je istoveten z: EN ISO 4307:2021
ICS:
11.100.10 Diagnostični preskusni In vitro diagnostic test
sistemi in vitro systems
SIST EN ISO 4307:2022 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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SIST EN ISO 4307:2022

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SIST EN ISO 4307:2022


EN ISO 4307
EUROPEAN STANDARD

NORME EUROPÉENNE

November 2021
EUROPÄISCHE NORM
ICS 11.100.10 Supersedes CEN/TS 17305:2019
English Version

Molecular in vitro diagnostic examinations - Specifications
for pre-examination processes for saliva - Isolated human
DNA (ISO 4307:2021)
Analyse de diagnostic moléculaire in vitro - Molekularanalytische in-vitro-diagnostische Verfahren
Spécifications relatives aux processus préanalytiques - Spezifikationen für präanalytische Prozesse für
pour la salive - ADN humain extrait (ISO 4307:2021) Speichel - Isolierte DNA (ISO 4307:2021)
This European Standard was approved by CEN on 18 October 2021.

CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this
European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references
concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN
member.

This European Standard exists in three official versions (English, French, German). A version in any other language made by
translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management
Centre has the same status as the official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and
United Kingdom.





EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2021 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 4307:2021 E
worldwide for CEN national Members.

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SIST EN ISO 4307:2022
EN ISO 4307:2021 (E)
Contents Page
European foreword . 3

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SIST EN ISO 4307:2022
EN ISO 4307:2021 (E)
European foreword
This document (EN ISO 4307:2021) has been prepared by Technical Committee ISO/TC 212 "Clinical
laboratory testing and in vitro diagnostic test systems" in collaboration with Technical Committee
CEN/TC 140 “In vitro diagnostic medical devices” the secretariat of which is held by DIN.
This European Standard shall be given the status of a national standard, either by publication of an
identical text or by endorsement, at the latest by May 2022, and conflicting national standards shall be
withdrawn at the latest by November 2024.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
This document supersedes CEN/TS 17305:2019.
Any feedback and questions on this document should be directed to the users’ national standards
body/national committee. A complete listing of these bodies can be found on the CEN website.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the
following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria,
Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland,
Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Republic of
North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the
United Kingdom.
Endorsement notice
The text of ISO 4307:2021 has been approved by CEN as EN ISO 4307:2021 without any modification.


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SIST EN ISO 4307:2022

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SIST EN ISO 4307:2022
INTERNATIONAL ISO
STANDARD 4307
First edition
2021-10
Molecular in vitro diagnostic
examinations — Specifications for
pre-examination processes for saliva
— Isolated human DNA
Analyse de diagnostic moléculaire in vitro — Spécifications relatives
aux processus préanalytiques pour la salive — ADN humain extrait
Reference number
ISO 4307:2021(E)
© ISO 2021

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SIST EN ISO 4307:2022
ISO 4307:2021(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2021
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on
the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address below
or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: +41 22 749 01 11
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
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SIST EN ISO 4307:2022
ISO 4307:2021(E)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2  Normative references . 1
3  Terms and definitions . 1
4 General considerations .4
5 Activities outside the laboratory.5
5.1 Specimen collection . 5
5.1.1 Information about the specimen donor/patient. 5
5.1.2 Selection of the saliva collection device by the laboratory . 5
5.1.3 Saliva specimen collection from the donor/patient and stabilization
procedures . 6
5.1.4 Information on the specimen and storage requirements at saliva collection
facility/site . 7
5.2 Transport requirements . 8
5.2.1 General . 8
5.2.2 Using saliva collection devices with DNA stabilizers . 8
5.2.3 Using saliva collection devices without DNA stabilizers . 8
6 Activities inside the laboratory . 9
6.1 Specimen reception . 9
6.2 Storage requirements . 9
6.3 Isolation of the saliva DNA . 9
6.3.1 General . 9
6.3.2 Using a commercial kit . 10
6.3.3 Using the laboratory’s own protocol . 10
6.4 Quantity and quality assessment of isolated DNA . 10
6.5 Storage of isolated saliva DNA . 11
6.5.1 General . 11
6.5.2 Saliva DNA isolated with commercially available kits . 11
6.5.3 Saliva DNA isolated with the laboratory’s own protocols . 11
Bibliography .12
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SIST EN ISO 4307:2022
ISO 4307:2021(E)
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 212, Clinical laboratory testing and in
vitro diagnostic test systems, in collaboration with the European Committee for Standardization (CEN)
Technical Committee CEN/TC 140, in vitro diagnostic medical devices, 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.
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SIST EN ISO 4307:2022
ISO 4307:2021(E)
Introduction
Molecular in vitro diagnostics has enabled a significant progress in medicine. Further progress is
expected by new technologies analysing profiles of nucleic acids, proteins, and metabolites in human
tissues and body fluids. However, the profiles of these molecules can change drastically during
specimen collection, transport, storage and processing thus making the outcome from diagnostics or
research unreliable or even impossible because the subsequent analytical assay will not determine the
situation in the patient but an artificial profile generated during the pre-examination process.
Genetic examination of DNA is commonly used in clinical practice. This includes, for example,
predisposition testing, pharmacogenomics and analysis of genetic disorders with the perspective used
in precision medicine. This is a fast-growing field in molecular diagnostics.
Saliva is increasingly used as a non-invasive alternative specimen to blood for the examination of
human DNA. Saliva naturally contains microorganisms and extraneous substances (e.g. food debris),
which make the composition of saliva more complex and unique among patients/donors. Dedicated
measures are therefore needed for informing and preparing patients/donors for the collection and
to check compliance with the instructions, in order to reduce the specimen variability. In contrast to
invasive specimen collection, saliva collection does not require trained and educated professionals or
dedicated facilities. By good instruction and verified collection device safety claims, saliva specimens
can be self-collected at home; however, home collection also contributes to high variability in specimen
quality. Similarly, medical laboratories/in vitro manufacturers need to be aware of specimen variability
when performing design verification and validation.
DNA in saliva can fragment or degrade after collection. In addition, bacteria present in the saliva
specimen can continue to grow, thus diluting the human DNA. DNases secreted by these bacteria can
also accelerate the DNA degradation. This can impact the sensitivity and reliability of DNA examination.
Standardization of the entire process from specimen collection to the DNA examination is needed to
minimize pre-examination impacts such as DNA degradation and fragmentation after saliva collection.
This document describes special measures which need to be taken to obtain good quality saliva
specimen/samples and isolated DNA therefrom for human DNA examination.
In this document, the following verbal forms are used:
— “shall” indicates a requirement;
— “should” indicates a recommendation;
— “may” indicates a permission;
— “can” indicates a possibility or a capability.
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SIST EN ISO 4307:2022
INTERNATIONAL STANDARD ISO 4307:2021(E)
Molecular in vitro diagnostic examinations —
Specifications for pre-examination processes for saliva —
Isolated human DNA
1 Scope
This document specifies requirements and recommendations on the handling, storage, processing and
documentation of saliva specimens intended for human DNA examination during the pre-examination
phase before a molecular examination is performed.
This document is applicable to molecular in vitro diagnostic examination including laboratory
developed tests performed by medical laboratories. It can also be used by laboratory customers, in
vitro diagnostics developers and manufacturers, biobanks, institutions and commercial organizations
performing biomedical research, and regulatory authorities.
Dedicated measures that need to be taken for saliva collected on absorbing material or by mouth
washes are not described in this document. Neither are measures for preserving and handling of native
saliva cell-free DNA, pathogens, and other bacterial or whole microbiome DNA in saliva described.
NOTE International, national or regional regulations or requirements can also apply to specific topics
covered in this document.
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 15190, Medical laboratories — Requirements for safety
3  Terms and definitions
For the purposes of this document, the terms and definitions given in ISO 15189 and the following apply.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at http:// www .electropedia .org/
3.1
ambient temperature
unregulated temperature of the surrounding air
[SOURCE: ISO 20184-1:2018, 3.2]
3.2
analyte
component represented in the name of a measurable quantity
[SOURCE: ISO 17511:2003, 3.2, modified — The examples were not taken over.]
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SIST EN ISO 4307:2022
ISO 4307:2021(E)
3.3
examination performance
analytical test performance
analytical performance
accuracy, precision, and sensitivity of a test to measure the analyte (3.2) of interest
Note 1 to entry: Other test performance characteristics such as robustness, repeatability can apply as well.
[SOURCE: ISO 20184-1:2018, 3.4]
3.4
DNA stabilizer
compound, solution or mixture that is designed to minimize degradation and fragmentation of DNA
(3.6)
[SOURCE: ISO 20186-2:2019, 3.5, modified — The term “genomic DNA in blood” has been replaced with
“DNA”.]
3.5
closed system
non-modifiable system provided by the vendor including all necessary components for the analysis (i.e.
hardware, software, procedures and reagents)
[SOURCE: ISO 20186-2:2019, 3.6]
3.6
DNA
deoxyribonucleic acid
polymer of deoxyribonucleotides occurring in a double-stranded (dsDNA) or single-stranded (ssDNA)
form
[SOURCE: ISO 22174:2005, 3.1.2]
3.7
examination
analytical test
set of operations having the object of determining the value or characteristics of a property
Note 1 to entry: Processes that start with the isolated measurand and include all kinds of parameter testing or
chemical manipulation for quantitative or qualitative examination.
[SOURCE: ISO 15189:2012, 3.7, modified — The term and definition is used here without the original
notes.]
3.8
examination provider
analytical test provider
entity that provides the specific analytical test
3.9
interfering substance
endogenous or exogenous substance (e.g. stabilization solution) that can be present in specimens and
that can alter an examination result
[SOURCE: ISO 20184-1:2018, 3.12]
3.10
microorganism
entity of microscopic size, encompassing bacteria, fungi and protozoa
[SOURCE: ISO 11139:2018, 3.176, modified — The term “viruses” was deleted from the definition.]
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SIST EN ISO 4307:2022
ISO 4307:2021(E)
3.11
pre-examination process
pre-analytical phase
pre-analytical workflow
process that starts, in chronological order, from the clinician’s request and include the examination
request, preparation and identification of the patient, collection of the primary sample(s) (3.12),
transportation to and within the analytical laboratory, isolation of analytes (3.2), and ends when the
analytical examination begins
Note 1 to entry: The pre-examination phase includes preparative processes that influence the outcome of the
intended examination.
[SOURCE: ISO 15189:2012, 3.15, modified — An additional term "pre-analytical workflow" was added,
and more detail was included.]
3.12
primary sample
specimen
discrete portion of a body fluid, breath, hair or tissue taken for examination, study or analysis of one or
more quantities or properties assumed to apply for the whole
[SOURCE: ISO 15189:2012, 3.16, modified — The term and definition is used here without the original
notes.]
3.13
proficiency testing
evaluation of participant performance against pre-established criteria by means of interlaboratory
comparisons
[SOURCE: ISO/IEC 17043:2010, 3.7, modified — Term and definition are used here without the original
notes.]
3.14
room temperature
temperature in the range of 18 °C to 25 °C, for the purpose of this document
Note 1 to entry: Local or national regulations can have different definitions.
[SOURCE: ISO 20186-2:2019, 3.22]
3.15
saliva
whole saliva
bio-fluid of the mouth composed mainly of secretion originating from the three major salivary glands
(parotids, submandibular and sublingual glands) and from salivary glands present in the oral cavity
3.16
saliva collection device
tube or other container in which the saliva (3.15) specimen (3.12) is collected
3.17
sample
one or more parts taken from a primary sample (3.12)
[SOURCE: ISO 15189:2012, 3.24, modified — The examples were not taken over.]
3.18
stability
ability of a specimen (3.12)/sample (3.17) material, when stored under specified conditions, to maintain
a defined property value within specified limits for a specified period of time
Note 1 to entry: The measurand constituent for the purpose of this document is isolated DNA (3.6).
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SIST EN ISO 4307:2022
ISO 4307:2021(E)
[SOURCE: ISO Guide 30:2015, 2.1.15, modified — Note 1 was not taken over. The following words
were replaced: “characteristic” by “ability”; “reference material” by “sample material”; “specified” by
“defined”.]
3.19
storage
prolonged interruption of the pre-analytical workflow (3.11) of a specimen (3.12) or sample (3.17) or
analyte (3.2) respectively, or of their derivatives, under appropriate conditions in order to preserve
their properties
Note 1 to entry: Long-term storage typically occurs in laboratory archives or in biobanks.
[SOURCE: ISO 20184-1:2018, 3.22, modified — Example in the definition was deleted and "specimen"
was added.]
3.20
validation
confirmation, through the provision of objective evidence, that the requirements for a specific intended
use or application have been fulfilled
Note 1 to entry: The word “validated” is used to designate the corresponding status.
[SOURCE: ISO 9000:2015, 3.8.13, modified — Note 1 and 3 were not taken over.]
3.21
verification
confirmation, through provision of objective evidence, that specified requirements have been fulfilled
Note 1 to entry: The word “verified” is used to designate the corresponding status.
Note 2 to entry: Confirmation can comprise activities such as:
— performing alternative calculations;
— comparing a new design specification with a similar proven design specification;
— undertaking tests and demonstrations; and
— reviewing documents prior to issue.
[SOURCE: ISO 9000:2015, 3.8.12, modified — Note 1 and Note 2 were not taken over.]
3.22
workflow
structured series of activities necessary to complete a task
[SOURCE: ISO 20184-1:2018, 3.26, modified — The word “structured” was added]
4 General considerations
For general statements on medical laboratory quality management systems and in particular on
primary sample collection, reception and handling (including avoidance of cross contaminations) see
ISO 15189 or ISO/IEC 17020. The requirements on laboratory equipment, reagents, and consumables
according to ISO 15189 shall be followed; ISO 15189 and ISO/IEC 17020 can also apply. For general
considerations on specimen collection, transport, receipt, handling, and storage, see ISO/TS 20658. For
biobanking, ISO 20387 can also apply.
All steps of a diagnostic workflow can influence the final examination result. Thus, the entire workflow,
including specimen/sample storage and transport conditions, and their impact on the stability of
biomolecules intended to be examined shall be specified, verified and validated for its intended use.
This includes the development of in vitro diagnostic (IVD) medical devices. The stability of the human
DNA should be investigated throughout the complete pre-examination process development. The
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SIST EN ISO 4307:2022
ISO 4307:2021(E)
verification of performance claims as well as the validation of the intended examination shall take into
account the variability of the saliva specimen's quality.
During the design and development of a saliva DNA based examination, a risk assessment shall be
performed (see also ISO 14971). Mitigation measures for eliminating or reducing identified risks shall
be established where required for ensuring the performance of the examination. This shall include the
pre-examination workflow steps.
Before or during the design of an examination, it should be investigated and ensured that the human
DNA quality parameters such as minimum DNA amount, size and purity required for the examination
is/are not compromised in a manner impacting the examination performance.
Safety requirements on specimen collection, transport and handling shall be in accordance with
relevant ISO standards such as ISO 15189 and ISO 15190.
During the whole pre-examination process, precautions shall be taken to avoid cross contamination
between different specimens/samples, for example by using single-use material whenever feasible or
appropriate cleaning procedures between processing of different specimens/samples.
For all pre-examination steps, the examination manufacturer's instructions shall be followed, if
provided.
Where, for justified reasons (e.g. unmet patient needs), a commercial product is not used in accordance
with the manufacturer's instructions, responsibility for its verification, validation, use and performance
lies with the laboratory.
The manufacturer's material safety data sheet should be considered before first use of any potentially
hazardous material (e.g. chemicals in stabilizers).
5 Activities outside the laboratory
5.1 Specimen collection
5.1.1  Information about the specimen donor/patient
The documentation shall include the identity of the specimen donor/patient, which can be in the form
of a code.
The documentation should include, but is not limited to:
a) the relevant health status of the primary sample donor/patient [e.g. healthy, disease type,
concomitant disease, demographics (e.g. age and sex)];
b) the information about medical treatment and special treatment prior to saliva collection (e.g.
anaesthetics, medications);
c) the type and the purpose of the examination requested;
d) the appropriate consent from the specimen donor/patient. See also ISO 15189.
5.1.2  Selection of the saliva collection device by the laboratory
The Saliva DNA examination manufacturer instructions should contain specifications on the saliva
collection device(s) to be used. Where the examination manufacture
...