EN ISO 20184-1:2018

Molecular in vitro diagnostic examinations - Specifications for pre-examination processes for frozen tissue - Part 1: Isolated RNA (ISO 20184-1:2018)

This document gives guidelines on the handling, documentation, storage and processing of frozen tissue specimens intended for RNA examination during the pre-examination phase before a molecular assay is performed.
This document is applicable to any molecular in vitro diagnostic examination performed by medical laboratories and molecular pathology laboratories that evaluate RNA extracted from frozen tissue. It is also intended to be used by laboratory customers, in vitro diagnostics developers and manufacturers, biobanks, institutions and commercial organisations performing biomedical research, and regulatory authorities.
Tissues that have undergone chemical stabilization pre-treatment before freezing are not covered in this document.
NOTE       International, national or regional regulations or requirements can also apply to specific topics covered in this document.

Molekularanalytische in-vitro-diagnostische Verfahren - Spezifikationen für präanalytische Prozesse für schockgefrorene Gewebeproben - Teil 1: Isolierte RNS (ISO 20184-1:2018)

Dieses Dokument dient als Leitfaden zur Handhabung, Dokumentation, Lagerung und Verarbeitung von gefrorenen Gewebeproben, die für die RNA Untersuchung während der präanalytischen Phase vor Beginn der molekularen Analyse vorgesehen sind.
Dieses Dokument ist anwendbar auf molekulare in vitro diagnostische Untersuchungen, die von medizinischen Laboratorien und Laboratorien der molekularen Pathologie durchgeführt werden, die aus gefrorenem Gewebe extrahierte RNA auswerten. Es hat außerdem den Zweck, von Kunden des Laboratoriums, Entwicklern und Herstellern im Bereich der In vitro Diagnostik, Biobanken, Institutionen und kommerziellen Organisationen, die biomedizinische Forschungen durchführen, und von Arzneimittel-agenturen verwendet zu werden.
Gewebe, die vor dem Gefriervorgang einer chemischen Vorbehandlung zur Stabilisierung unterzogen wurden, sind nicht durch dieses Dokument abgedeckt.
ANMERKUNG   Internationale, nationale oder regionale Regelungen bzw. Anforderungen können ebenfalls für bestimmte Themen in diesem Dokument gelten.

Analyses de diagnostic moléculaire in vitro - Spécifications relatives aux processus préanalytiques pour les tissus congelés - Partie 1: ARN extrait (ISO 20184-1:2018)

Le présent document fournit des lignes directrices pour la manipulation, la documentation, le stockage et le traitement de prélèvements de tissus congelés destinés à l'analyse de l'ARN durant la phase préanalytique précédant la réalisation d'un essai moléculaire.
Le présent document s'applique aux analyses de diagnostic moléculaire in vitro réalisées par des laboratoires de biologie médicale et des laboratoires de pathologie moléculaire qui évaluent l'ARN extrait de tissus congelés. Il est également destiné à être utilisé par des clients de laboratoires, des développeurs et fabricants de l'industrie du diagnostic in vitro, ainsi que par des biobanques, des institutions et des organismes commerciaux spécialisés en recherche biomédicale, de même que des autorités de réglementation.
Le cas des tissus ayant subi un prétraitement de stabilisation chimique avant la congélation n'est pas couvert par le présent document.
NOTE       Des réglementations ou exigences internationales, nationales ou régionales peuvent également s'appliquer à des sujets spécifiques traités dans le présent document.

Molekularne diagnostične preiskave in vitro - Specifikacije za predpreiskovalne procese za zamrznjena tkiva - 1. del: Izolirani RNK (ISO 20184-1:2018)

Ta mednarodni standard vsebuje priporočila za obravnavo, dokumentiranje, shranjevanje in obdelavo vzorcev zamrznjenih tkiv, namenjenih za analizo RNK med predpreiskovalno fazo, preden se izvede molekularni preskus. Ta mednarodni standard se uporablja za molekularne diagnostične preiskave in vitro, vključno z laboratorijsko razvitimi preskusi, ki jih izvajajo v medicinskih laboratorijih.  Uporabljali naj bi ga tudi uporabniki laboratorijev, razvijalci in proizvajalci diagnostike in vitro, nanaša pa se tudi na institucije in komercialne organizacije, ki izvajajo biomedicinske raziskave, biobanke ter regulativne organe. Profili RNK v tkivih se lahko močno spremenijo pred zbiranjem in po njem ter se različno spremenijo pri tkivih različnih darovalcev/bolnikov. Zato je nujno treba sprejeti posebne ukrepe, da se zmanjšajo opisane spremembe profila v tkivu za nadaljnje preiskave RNA. Tkiva, ki so pred zamrzovanjem prestala predobdelavo za kemično stabilizacijo, niso zajeta v tem dokumentu. OPOMBA:   Za določene teme, ki so zajete v tem mednarodnem standardu, lahko veljajo tudi mednarodni, nacionalni ali regionalni predpisi ali zahteve.

General Information

• Status: Published
• Publication Date: 18-Dec-2018
• Withdrawal Date: 29-Jun-2019
• ICS: 11.100.10 - In vitro diagnostic test systems
• Technical Committee: CEN/TC 140 - In vitro diagnostic systems
• Drafting Committee: CEN/TC 140/WG 3 - Quality management in the medical laboratory
• Current Stage: 6060 - Definitive text made available (DAV) - Publishing
• Start Date: 19-Dec-2018
• Due Date: 11-Aug-2018
• Completion Date: 19-Dec-2018
• Ref Project: SIST EN ISO 20184-1:2019 - Molecular in vitro diagnostic examinations - Specifications for pre-examination processes for frozen tissue - Part 1: Isolated RNA (ISO 20184-1:2018)

Relations

• Replaces: CEN/TS 16826-1:2015 - Molecular in vitro diagnostic examinations - Specifications for pre-examination processes for snap frozen tissue - Part 1: Isolated RNA
• Effective Date: 05-Sep-2018

Overview

EN ISO 20184-1:2018 (ISO 20184-1:2018) - "Molecular in vitro diagnostic examinations - Specifications for pre‑examination processes for frozen tissue - Part 1: Isolated RNA" provides standardized guidance for the pre‑examination phase of molecular IVD testing when RNA is extracted from frozen tissue. The standard focuses on minimizing changes to RNA profiles that can occur during collection, transport, handling, freezing, storage and initial processing so that downstream molecular assays accurately reflect the in‑vivo state.

Key topics and requirements

• Scope and applicability: Applies to medical and molecular pathology laboratories, biobanks, IVD developers/manufacturers, research organisations and regulatory authorities involved with RNA extracted from frozen tissue. Chemically stabilized tissues (pre‑treatment before freezing) are excluded.
• Pre‑examination controls: Emphasizes documentation and traceability for specimen donor/patient information and specimen details during the pre‑examination phase.
• Specimen collection and transport: Guidance on collection best practices, required information, preparations for transport and handling during transport to control pre‑analytical variability.
• Laboratory reception and evaluation: Reception documentation, pathological evaluation and selection of sample(s) for RNA analysis.
• Freezing and storage: Recommendations for freezing workflows and storage requirements for frozen tissues to preserve RNA integrity.
• RNA isolation: General requirements, recommendations for using commercial kits or laboratory‑developed protocols, and considerations to preserve RNA quality.
• RNA quality and quantity assessment: Requirements for assessing isolated RNA prior to molecular assays.
• Storage of isolated RNA: Guidance on post‑isolation storage conditions and record keeping.
• Informative annex: Includes impact of pre‑examination variables (example: liver tissue) on RNA profiles to illustrate practical consequences of non‑standardized handling.

Note: The standard uses prescriptive language - “shall” for requirements and “should” for recommendations - and refers to ISO 15189 for quality and competence needs.

Practical applications

• Ensures RNA integrity and reliable molecular diagnostic results for assays such as gene expression profiling, RNA sequencing and RNA‑based biomarker tests.
• Helps laboratories implement standardized sample handling, documentation and storage workflows to reduce pre‑analytical variability.
• Guides biobanks and clinical sites on specimen collection, labeling and transport procedures to maintain sample value for future RNA analyses.
• Supports IVD developers and manufacturers in designing kits and instructions that align with recognized pre‑analytical requirements.
• Aids regulatory bodies and quality managers in assessing laboratory conformity and traceability related to pre‑examination processes.

Related standards

• ISO 15189 (Medical laboratories - Requirements for quality and competence) - cited normative reference.
• Other parts of the ISO 20184 series (see ISO catalogue) covering complementary specimen types or analytes.

Keywords: EN ISO 20184-1:2018, ISO 20184-1, pre‑examination, frozen tissue, isolated RNA, molecular in vitro diagnostic, RNA integrity, sample handling, biobanks, molecular pathology.

Standards Content (Sample)

SLOVENSKI STANDARD
01-marec-2019
1DGRPHãþD
SIST-TS CEN/TS 16826-1:2015
0ROHNXODUQHGLDJQRVWLþQHSUHLVNDYHLQYLWUR6SHFLILNDFLMH]DSUHGSUHLVNRYDOQH
SURFHVH]D]DPU]QMHQDWNLYDGHO,]ROLUDQL51.,62
Molecular in vitro diagnostic examinations - Specifications for pre-examination processes
for frozen tissue - Part 1: Isolated RNA (ISO 20184-1:2018)
Molekularanalytische in-vitro-diagnostische Verfahren - Spezifikationen für
präanalytische Prozesse für schockgefrorene Gewebeproben - Teil 1: Isolierte RNS (ISO
20184-1:2018)
Examens de diagnostic moléculaire in vitro - Spécifications pour les processus
d'examens préliminaires des tissus congelés - Partie 1: ARN isolé (ISO 20184-1:2018)
Ta slovenski standard je istoveten z: EN ISO 20184-1:2018
ICS:
11.100.10 'LDJQRVWLþQLSUHVNXVQL In vitro diagnostic test
VLVWHPLLQYLWUR systems
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EN ISO 20184-1
EUROPEAN STANDARD
NORME EUROPÉENNE
December 2018
EUROPÄISCHE NORM
ICS 11.100.10 Supersedes CEN/TS 16826-1:2015
English Version
Molecular in vitro diagnostic examinations - Specifications
for pre-examination processes for frozen tissue - Part 1:
Isolated RNA (ISO 20184-1:2018)
Analyses de diagnostic moléculaire in vitro - Molekularanalytische in-vitro-diagnostische Verfahren
Spécifications relatives aux processus préanalytiques - Spezifikationen für präanalytische Prozesse für
pour les tissus congelés - Partie 1: ARN extrait (ISO schockgefrorene Gewebeproben - Teil 1: Isolierte RNS
20184-1:2018) (ISO 20184-1:2018)
This European Standard was approved by CEN on 30 November 2018.

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, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, 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
© 2018 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 20184-1:2018 E
worldwide for CEN national Members.

Contents Page
European foreword . 3

European foreword
This document (EN ISO 20184-1:2018) 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 June 2019, and conflicting national standards shall be
withdrawn at the latest by June 2019.
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 16826-1:2015.
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, Former Yugoslav Republic of Macedonia,
France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta,
Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,
Turkey and the United Kingdom.
Endorsement notice
The text of ISO 20184-1:2018 has been approved by CEN as EN ISO 20184-1:2018 without any
modification.
INTERNATIONAL ISO
STANDARD 20184-1
First edition
2018-11
Molecular in vitro diagnostic
examinations — Specifications for
pre-examination processes for frozen
tissue —
Part 1:
Isolated RNA
Analyses de diagnostic moléculaire in vitro — Spécifications relatives
aux processus préanalytiques pour les tissus congelés —
Partie 1: ARN extrait
Reference number
ISO 20184-1:2018(E)
©
ISO 2018
ISO 20184-1:2018(E)
© ISO 2018
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: [phone]
Fax: [phone]
Email: [e-mail]
Website: www.iso.org
Published in Switzerland
ii © ISO 2018 – All rights reserved

ISO 20184-1:2018(E)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2  Normative references . 1
3  Terms and definitions . 1
4 General considerations . 5
5 Outside the laboratory . 5
5.1 Specimen collection . 5
5.1.1 General. 5
5.1.2 Information about the specimen donor/patient . 6
5.1.3 Information about the specimen . 6
5.1.4 Specimen processing . 6
5.2 Fresh tissue transport requirements . 7
5.2.1 General. 7
5.2.2 Preparations for the transport . 7
5.2.3 During transport . 7
6 Inside the laboratory . 7
6.1 Information about the reception of the specimen . 7
6.2 Evaluation of the pathology of the specimen and selection of the sample(s) . 8
6.3 Freezing of the specimen or sample(s) . 8
6.4 Storage requirements .10
6.5 Isolation of RNA .11
6.5.1 General.11
6.5.2 Requirements and recommendations .11
6.5.3 Using commercial kits .12
6.5.4 Using the laboratories own protocols .12
6.6 Quantity and quality assessment of isolated RNA .12
6.7 Storage of isolated RNA .12
6.7.1 General.12
6.7.2 Using commercially available kits for RNA isolation .13
6.7.3 Using the laboratory's own protocols for RNA isolation .13
Annex A (informative) Impact of pre-examination variables on RNA profiles obtained from
frozen liver tissue samples collected during and after routine surgery .14
Bibliography .18
ISO 20184-1:2018(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.
A list of all parts in the ISO 20184 can be found on the ISO website.
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 © ISO 2018 – All rights reserved

ISO 20184-1:2018(E)
Introduction
Molecular in vitro diagnostics, including molecular pathology, has enabled a significant progress in
medicine. Further progress is expected with new technologies analysing nucleic acids, proteins, and
metabolites in human tissues and body fluids. However, the profiles and/or integrity 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
examination assay will not determine the situation in the patient but an artificial profile generated
during the pre-examination process.
Therefore, a standardization of the entire process from specimen collection to the RNA examination is
needed. Studies have been undertaken to determine the important influencing factors. This document
draws upon such work to codify and standardize the steps for frozen tissue with regard to RNA
examination in what is referred to as the pre-examination phase.
RNA profiles in tissues can change drastically before, during and after collection (due to e.g. gene
induction or gene down regulation). RNA species can change differently in different donor's patients’
tissues.
Therefore, it is essential to take special measures to minimize the described RNA profile changes and
modifications within the tissue for subsequent 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.
INTERNATIONAL STANDARD ISO 20184-1:2018(E)
Molecular in vitro diagnostic examinations —
Specifications for pre-examination processes for frozen
tissue —
Part 1:
Isolated RNA
1 Scope
This document gives guidelines on the handling, documentation, storage and processing of frozen
tissue specimens intended for RNA examination during the pre-examination phase before a molecular
assay is performed.
This document is applicable to any molecular in vitro diagnostic examination performed by medical
laboratories and molecular pathology laboratories that evaluate RNA extracted from frozen tissue. It is
also intended to be used by laboratory customers, in vitro diagnostics developers and manufacturers,
biobanks, institutions and commercial organisations performing biomedical research, and regulatory
authorities.
Tissues that have undergone chemical stabilization pre-treatment before freezing are not covered in
this document.
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 15189:2012, Medical laboratories — Requirements for quality and competence
3  Terms and definitions
For the purposes of this document, the terms and definitions given in ISO 15189 and the following
terms and definitions 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
aliquot
portion of a larger amount of homogenous material, assumed to be taken with negligible sampling error
Note 1 to entry: The term is usually applied to fluids. Tissues are heterogeneous and therefore cannot be
aliquoted.
Note 2 to entry: The definition is derived from References [22], [23] and [24].
ISO 20184-1:2018(E)
3.2
ambient temperature
unregulated temperature of the surrounding air
3.3
analyte
component represented in the name of a measurable quantity
[SOURCE: ISO 17511:2003, 3.2]
3.4
analytical test performance
accuracy, precision, and sensitivity of a test to measure the analyte of interest
Note 1 to entry: Other test performance characteristics such as robustness, repeatability can apply as well.
3.5
cold ischemia
condition after removal of the tissue from the body until stabilization or fixation
3.6
diagnosis
identification of a health or disease state from its signs and/or symptoms, where the diagnostic process
can involve examinations and tests for classification of an individual's condition into separate and
distinct categories or subclasses that allow medical decisions about treatment and prognosis to be made
3.7
DNA
deoxyribonucleic acid
polymer of deoxyribonucleotides occurring in a double-stranded (dsDNA) or single-stranded
(ssDNA) form
[SOURCE: ISO 22174:2005, 3.1.2]
3.8
DNase
deoxyribonuclease
enzyme that catalyzes the degradation of DNA into smaller components
3.9
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 analyte and include all kinds of parameter testing or
chemical manipulation for quantitative or qualitative examination.
[SOURCE: ISO 15189:2012, 3.7, modified — Notes to entry 1 to 3 have been removed, Note 1 to entry has
been added and “analytical test” has been added as a preferred term.]
3.10
grossing
gross examination
inspection of pathology specimens with the bare eye to obtain diagnostic information, while being
processed for further microscopic examination
3.11
homogeneous
uniform in structure and composition
2 © ISO 2018 – All rights reserved

ISO 20184-1:2018(E)
3.12
interfering substances
endogenous substances of a specimen/sample or exogenous substances (e.g. stabilization solution) that
can alter an examination result
3.13
pre-examination processes
preanalytical phase
preanalytical workflow
processes that start, 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), transportation
to and within the medical or pathology laboratory, isolation of analytes, and end 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 — “pre-analytical workflow” has been added as a preferred
term, Note 1 to entry has been added and the definition has been extended.]
3.14
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 — Notes to entry 1 to 3 have been removed.]
3.15
proficiency test
evaluation of participant performance against pre-established criteria by means of inter-laboratory
comparisons
[SOURCE: ISO 17043:2010, 3.7, modified — Notes to entry 1 and 2 have been removed.]
3.16
RNA profile
amounts of the individual RNA molecules that are present in a sample and that can be measured in the
absence of any losses, inhibition and interference
3.17
RNA
ribonucleic acid
polymer of ribonucleotides occurring in a double-stranded or single-stranded form
[SOURCE: ISO 22174:2005, 3.1.3]
3.18
RNase
ribonuclease
enzyme that catalyses the degradation of RNA into smaller components
3.19
room temperature
for the purposes of this document, temperature in the range of 18 °C to 25 °C
Note 1 to entry: Local or national regulations can have different definitions.
ISO 20184-1:2018(E)
3.20
sample
one or more parts taken from a primary sample
[SOURCE: ISO 15189:2012, 3.24, modified — EXAMPLE has been removed.]
3.21
stability
characteristic of a sample material, when stored under specified conditions, to maintain a specified
property value within specified limits for a specified period of time
Note 1 to entry: The analyte for the purpose of this document is isolated RNA.
[SOURCE: ISO Guide 30:2015, 2.1.15, modified — “reference material” has been replaced by “sample
material”, “characteristic” has been replaced by “ability” and Note 1 to entry has been changed.]
3.22
storage
prolonged interruption of the pre-analytical workflow of a sample or analyte respectively, or of their
derivatives e.g., stained sections or tissue blocks, under appropriate conditions in order to preserve
their properties
Note 1 to entry: Long-term storage typically occurs in laboratory archives or in biobanks.
3.23
validation
confirmation, throughout the provision of objective evidence, that the requirements for a specific
intended use or application have been fulfilled
Note 1 to entry: The term “validated” is used to designate the corresponding status.
[SOURCE: ISO 9000:2015, 3.8.13, modified — Notes to entry 1 and 3 have been removed.]
3.24
verification
confirmation, through provision of objective evidence, that specified requirements have been fulfilled
Note 1 to entry: The term “verified” is used to designate the corresponding status.
[SOURCE: ISO 9000:2015, 3.8.12, modified — Note 1 and Note 2 where not taken over.]
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;
— reviewing documents prior to issue.
3.25
warm ischemia
condition before the tissue is removed from the body, but where it is deprived of its normal blood supply
3.26
workflow
series of activities necessary to complete a task
4 © ISO 2018 – All rights reserved

ISO 20184-1:2018(E)
4 General considerations
For general statements on medical laboratory quality management systems and in particular on
specimen collection, reception and handling (including avoidance of cross contaminations) see
ISO 15189:2012, 4.2, 5.4.4, 5.4.6 or ISO/IEC 17020:2012, Clause 8 and 7.2. The requirements on
laboratory equipment, reagents, and consumables in accordance with ISO 15189:2012, 5.3 shall be
followed; ISO 15189:2012, 5.5.1.2 and 5.5.1.3, and ISO/IEC 17020:2012, 6.2 can also apply.
All steps of a diagnostic workflow can influence the final analytical test result. Thus, the entire
workflow including biomolecule stability and sample storage conditions shall be verified and validated.
Workflow steps which cannot always be controlled (e.g. warm ischemia) shall be documented. A risk
assessment of non-controllable workflow steps including their potential impact on the analytical test
performance shall be performed and mitigation measures shall be established to enable the required
analytical test performance.
Before or during the design of an analytical test, it should therefore be investigated and assured that the
RNA profile(s) intended to be analysed is/are not compromised in a manner impacting the analytical
test performance (e.g. by performing a time course experiment or study; see also Annex A).
Before tissues are stabilized by freezing, the RNA profile(s) can change e.g. by gene induction, gene
down regulation and RNA degradation. These effects depend on the warm and cold ischemia duration
and the ambient temperature before freezing. In addition, the described effects can vary in different
donors'/patients' tissues.
Generally, the longer the duration of warm and cold ischemia and the higher the ambient temperature
before freezing the tissue specimen, the higher is the risk that changes in the RNA profile can occur.
NOTE Intraoperative warm ischemia can result in more pronounced changes of RNA profiles than during
[7][8]
postoperative cold ischemia . RNA profiles can also vary, depending on the origin and type of tissue, the
underlying disease, the surgical procedure, the drug regimen, and drugs administered for anaesthesia or
treatment of concomitant disease and on the different environmental conditions after the tissue removal from
the body.
As warm ischemia cannot be easily standardized, its duration shall be documented. When it is not
possible to avoid cold ischemia, duration shall be documented and temperatures of the specimen
container's surroundings should be documented. Where the specimen is transported to another facility
for freezing, the transport duration shall be documented and the ambient conditions should also be
documented.
Safety instructions on transport and handling shall be considered (see ISO 15189:2012, 5.2.3 and 5.4.5
and ISO 15190).
During the whole pre-examination process precautions shall be taken to avoid cross contamination
between different specimens/samples, e.g. by using single-use material whenever feasible or
appropriate cleaning procedures between processing of different specimens/samples.
If a commercial product is not used in accordance with the manufacturer’s instructions, responsibility
for its use and performance lies with the user.
5 Outside the laboratory
5.1 Specimen collection
5.1.1 General
For the collection of the specimen, the requirements (e.g. disease condition, specimen size) for the
intended molecular examination (see also Clause 6) should be considered.
See also ISO 15189:2012, 5.4.4.
ISO 20184-1:2018(E)
5.1.2  Information about the specimen donor/patient
The documentation shall include the ID 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 specimen donor/patient (e.g. healthy, disease type, concomitant
disease, demographics [e.g. age and gender]);
b) the information about routine medical treatment and special treatment prior to tissue collection
(e.g. anaesthetics, medications, surgical or diagnostic procedures);
c) the appropriate consent from the specimen donor/patient.
5.1.3  Information about the specimen
The documentation shall include, but is not limited to:
a) the start of ischemia within the body (warm ischemia) by documentation of the ischemia-relevant
vessel ligation/clamping time point (usually arterial clamping time);
NOTE Not needed where small tissue biopsy resection for freezing is performed.
b) the time and date when tissue is removed from the body and the method of removal (e.g. core-
needle biopsy, resection, biopsy device used for the collection);
c) the description of tissue type and origin, tissue condition (e.g. diseased, unaffected by the disease),
including references to any marking applied in or outside the operating theatre made by surgeon,
radiologist or pathologist.
If the freezing of the tissue is performed outside the laboratory, the documentation of steps described
under 6.2, where pathology evaluation is required, and 6.3 has to be performed.
The documentation should also include the ID of the responsible person collecting the specimen.
5.1.4 Specimen processing
Tissues that need to be frozen for diagnostic purposes can originate from a large tissue specimen or
can be a small tissue specimen like biopsies e.g. taken by endoscopy or taken from patients during a
surgical procedure where fast frozen section diagnosis is required.
Post-mortem tissues can be frozen for diagnostic purposes. However, preservation of RNA is dependent
on the time interval between death and autopsy and the temperature of storage of the body after death.
Any additions or modifications to the specimen after removal from the body (e.g. labelling for the
orientation of the specimen [e.g. ink-marking, stitches, incision(s)]) shall be documented.
Where a pathology diagnosis is required on the specimen, sampling shall be performed by or under
supervision or guidance of a medically qualified (e.g. board certified) pathologist (see 6.2).
Where the specimen was removed without the requirement of pathology diagnosis, the evaluation,
selection and documentation of specimens may be done by other qualified persons than pathologists.
The freezing of the specimen or samples taken from the specimen can be performed outside the
laboratory or inside the laboratory.
Cold ischemia can influence the RNA profile; therefore direct freezing should be preferred.
a) In case the specimen or sample is frozen outside the laboratory, proceed with 6.2 without delay.
6 © ISO 2018 – All rights reserved

ISO 20184-1:2018(E)
b) In case the specimen or sample is frozen inside the laboratory, fresh tissue specimens need to be
transported to the laboratory. The steps described under 5.2 for fresh tissue transport shall be
performed without delay.
5.2 Fresh tissue transport requirements
5.2.1 Gene
...