Molecular in-vitro diagnostic examinations - Specifications for pre-examination processes for venous whole blood - Part 3: Isolated circulating cell free DNA from plasma (ISO 20186-3:2019)

This International Standard recommends the handling, documentation, storage and processing of venous whole blood specimens intended for circulating cell free DNA (ccfDNA) examination during the pre-examination phase before a molecular assay is performed. This International Standard covers specimens collected in venous whole blood collection tubes. This International Standard is applicable to molecular in vitro diagnostic examinations including laboratory developed tests performed by medical laboratories. It is also intended to be used by laboratory customers, in vitro diagnostics developers and manufacturers, but also pertains institutions and commercial organizations performing biomedical research, biobanks, and regulatory authorities.
CcfDNA profiles can change significantly after blood collection from the donor (e.g., release of genomic DNA from white blood cells, ccfDNA fragmentation and ccfDNA quantity change). Therefore, special measures have to be taken to secure good quality blood samples for ccfDNA examination and storage.
Different dedicated measures need to be taken for preserving blood genomic DNA, which are not described in this International Standard. Blood genomic DNA is covered in ISO 20185-2, Molecular in vitro diagnostic examinations — specifications for pre-examination processes for venous whole blood — Part 2: Isolated genomic DNA.
NOTE         CcfDNA obtained from blood by the procedures suggested in this document can contain DNA present in exosomes.
Pathogen DNA present in blood is not covered by this International Standard.
Different dedicated measures need to be taken for preserving DNA in circulating exosomes, which are not described in this International Standard.
NOTE   International, national or regional regulations or requirements may also apply to specific topics covered in this International Standard.

Molekularanalytische in-vitro-diagnostische Verfahren - Spezifikationen für präanalytische Prozesse für venöse Vollblutproben - Teil 3: Aus Plasma isolierte zirkulierende zellfreie DNS (ISO 20186-3:2019)

Dieses Dokument enthält Empfehlungen und Anforderungen für die Handhabung, Lagerung, Verarbeitung und Dokumentation von venösen Vollblutproben, die für die Untersuchung zirkulierender zellfreier DNA (ccfDNA) vorgesehen sind, während der präanalytischen Phase vor Beginn einer analytischen Prüfung. Dieses Dokument behandelt Untersuchungsmaterial, das in Blutentnahmeröhrchen für venöses Vollblut entnommen wurde.
Dieses Dokument gilt für alle molekularen in vitro diagnostischen Untersuchungen, die in medizinischen Laboren durchgeführt werden. Es ist außerdem auch für die Verwendung durch Labor Kunden, Entwickler und Hersteller von In vitro Diagnostika, Biobanken, Einrichtungen und kommerzielle Organisationen, die in der biomedizinischen Forschung tätig sind, sowie durch Aufsichtsbehörden vorgesehen.
Zur Stabilisierung genomischer DNA aus Blut werden andere spezielle Maßnahmen getroffen, die nicht in diesem Dokument beschrieben sind. Genomische DNA aus Blut wird in ISO 20186 2 behandelt.
Zur Erhaltung der DNA in zirkulierenden Exosomen werden andere zweckbestimmte Maßnahmen ergriffen, die in diesem Dokument nicht beschrieben werden.
ANMERKUNG   ccfDNA, die mit den in diesem Dokument vorgeschlagenen Verfahren aus Blut gewonnen wurde, kann DNA enthalten, die ursprünglich in Exosomen vorlag [8], [9].
Nicht durch dieses Dokument abgedeckt ist DNA, die in Krankheitserregern im Blut vorhanden ist.

Tests de diagnostic moléculaire in vitro - Spécifications relatives aux processus pré-analytiques pour le sang - ARN cellulaire - Partie 3: ADN libre circulant extrait du plasma (ISO 20186-3:2019)

Le présent document fournit des recommandations et des exigences sur la manipulation, le stockage, le traitement et la documentation des prélèvements de sang total veineux destinés à l'analyse de l'ADN libre circulant (ADNlc) durant la phase préanalytique précédant la réalisation d'un essai analytique. Le présent document concerne les échantillons primaires prélevés dans des tubes de prélèvement de sang total veineux.
Le présent document s'applique aux analyses de diagnostic moléculaire in vitro réalisées par des laboratoires de biologie médicale. 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.
Des mesures spécifiques différentes, non décrites dans le présent document, sont prises pour stabiliser l'ADN génomique sanguin. L'ADN génomique sanguin est couvert par l'ISO 20186-2.
Des mesures spécifiques différentes, non décrites dans le présent document, sont prises pour préserver l'ADN des exosomes circulants.
NOTE L'ADNlc extrait du sang à l'aide des protocoles cités dans le présent document est susceptible de contenir de l'ADN présent à l'origine dans les exosomes[8][9].
L'ADN des pathogènes présents dans le sang n'est pas couvert par le présent document.

Molekularne diagnostične preiskave in vitro - Specifikacije za predpreiskovalne procese za vensko polno kri - 3. del: Iz plazme izolirana cirkulirajoča brezcelična DNK (ISO 20186-3:2019)

Ta mednarodni standard vsebuje priporočila za obravnavo, dokumentiranje, shranjevanje in obdelavo vzorcev venske polne krvi, namenjenih za analizo cirkulirajoče brezcelične DNK (ccfDNA) med predpreiskovalno fazo, preden se izvede molekularni preskus. Ta mednarodni standard zajema vzorce, ki so zbrani s cevkami za zbiranje venske polne krvi. 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, navezuje pa se tudi na institucije in komercialne organizacije, ki izvajajo biomedicinske raziskave, biobanke ter regulativne organe. Profili cirkulirajoče brezcelične DNK se lahko znatno spremenijo po odvzemu krvi od darovalca (npr. sprostitev genomske DNK iz belih krvnih celic, razpad ali sprememba količine cirkulirajoče brezcelične DNK). Zato je treba sprejeti posebne ukrepe za pridobivanje vzorcev krvi dobre kakovosti za analizo in shrambo cirkulirajoče brezcelične DNK. Za ohranjanje krvne genomske DNK je treba uporabiti drugačne namenske ukrepe, ki niso opisani v tem mednarodnem standardu. Krvna genomska DNK je opisana v standardu ISO 20185-2, Molekularne diagnostične preiskave in vitro – specifikacije za predpreiskovalne procese za vensko polno kri - 2. del: Iz genoma izolirana DNK OPOMBA: cirkulirajoča brezcelična DNK iz krvi, pridobljena s postopki, ki jih predlaga ta dokument, lahko vsebuje DNK iz eksosomov. Patogena DNK v krvi ni zajeta v tem mednarodnem standardu. Za ohranjanje DNK v cirkulirajočih eksosomih je treba uporabiti drugačne namenske ukrepe, ki niso opisani v tem mednarodnem standardu. 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
Public Enquiry End Date
19-Mar-2018
Publication Date
10-Dec-2019
Technical Committee
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
20-Nov-2019
Due Date
25-Jan-2020
Completion Date
11-Dec-2019

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SLOVENSKI STANDARD
SIST EN ISO 20186-3:2020
01-januar-2020
Nadomešča:
SIST-TS CEN/TS 16835-3:2015
Molekularne diagnostične preiskave in vitro - Specifikacije za predpreiskovalne

procese za vensko polno kri - 3. del: Iz plazme izolirana cirkulirajoča brezcelična

DNK (ISO 20186-3:2019)

Molecular in-vitro diagnostic examinations - Specifications for pre-examination processes

for venous whole blood - Part 3: Isolated circulating cell free DNA from plasma (ISO

20186-3:2019)
Molekularanalytische in-vitro-diagnostische Verfahren - Spezifikationen für
präanalytische Prozesse für venöse Vollblutproben - Teil 3: Aus Plasma isolierte
zirkulierende zellfreie DNS (ISO 20186-3:2019)

Tests de diagnostic moléculaire in vitro - Spécifications relatives aux processus pré-

analytiques pour le sang - ARN cellulaire - Partie 3: ADN libre circulant extrait du plasma

(ISO 20186-3:2019)
Ta slovenski standard je istoveten z: EN ISO 20186-3:2019
ICS:
11.100.10 Diagnostični preskusni In vitro diagnostic test
sistemi in vitro systems
SIST EN ISO 20186-3:2020 en

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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SIST EN ISO 20186-3:2020
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SIST EN ISO 20186-3:2020
EN ISO 20186-3
EUROPEAN STANDARD
NORME EUROPÉENNE
October 2019
EUROPÄISCHE NORM
ICS 11.100.10
English Version
Molecular in-vitro diagnostic examinations - Specifications
for pre-examination processes for venous whole blood -
Part 3: Isolated circulating cell free DNA from plasma (ISO
20186-3:2019)

Analyses de diagnostic moléculaire in vitro - Molekularanalytische in-vitro-diagnostische Verfahren

Spécifications relatives aux processus préanalytiques - Spezifikationen für präanalytische Prozesse für

pour le sang total veineux - Partie 3: ADN libre venöse Vollblutproben - Teil 3: Aus Plasma isolierte

circulant extrait du plasma (ISO 20186-3:2019) zirkulierende zellfreie DNA (ISO 20186-3:2019)

This European Standard was approved by CEN on 14 September 2019.

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

© 2019 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 20186-3:2019 E

worldwide for CEN national Members.
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SIST EN ISO 20186-3:2020
EN ISO 20186-3:2019 (E)
Contents Page

European foreword ....................................................................................................................................................... 3

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SIST EN ISO 20186-3:2020
EN ISO 20186-3:2019 (E)
European foreword

This document (EN ISO 20186-3:2019) 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 April 2020, and conflicting national standards shall be

withdrawn at the latest by October 2022.

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 16835-3:2015.

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, 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 20186-3:2019 has been approved by CEN as EN ISO 20186-3:2019 without any

modification.
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SIST EN ISO 20186-3:2020
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SIST EN ISO 20186-3:2020
INTERNATIONAL ISO
STANDARD 20186-3
First edition
2019-09
Molecular in vitro diagnostic
examinations — Specifications for
pre-examination processes for venous
whole blood —
Part 3:
Isolated circulating cell free DNA
from plasma
Analyses de diagnostic moléculaire in vitro — Spécifications relatives
aux processus préanalytiques pour le sang total veineux —
Partie 3: ADN libre circulant extrait du plasma
Reference number
ISO 20186-3:2019(E)
ISO 2019
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SIST EN ISO 20186-3:2020
ISO 20186-3:2019(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2019

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
Fax: +41 22 749 09 47
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii © ISO 2019 – All rights reserved
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SIST EN ISO 20186-3:2020
ISO 20186-3:2019(E)
Contents Page

Foreword ........................................................................................................................................................................................................................................iv

Introduction ..................................................................................................................................................................................................................................v

1 Scope ................................................................................................................................................................................................................................. 1

2  Normative references ...................................................................................................................................................................................... 1

3  Terms and definitions ..................................................................................................................................................................................... 1

4 General consideration .................................................................................................................................................................................... 5

5 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 venous whole blood collection tube by the laboratory .......................... 6

5.1.3 Venous whole blood collection from the donor/patient and stabilization

procedures ............................................................................................................................................................................ 6

5.1.4 Information about the specimen and storage requirements at the blood

collection facility .............................................................................................................................................................. 7

5.2 Transport requirements ................................................................................................................................................................. 7

6 Inside the laboratory ....................................................................................................................................................................................... 8

6.1 Specimen reception ............................................................................................................................................................................ 8

6.2 Storage requirements for blood specimens ................................................................................................................... 8

6.3 Plasma preparation ............................................................................................................................................................................. 9

6.4 Storage requirements for plasma samples ..................................................................................................................... 9

6.5 Isolation of the ccfDNA ..................................................................................................................................................................10

6.5.1 General...................................................................................................................................................................................10

6.5.2 Using blood collection tubes with stabilizers ......................................................................................10

6.5.3 Using blood collection tubes without stabilizers ..............................................................................11

6.6 Quantity and quality assessment of isolated ccfDNA .........................................................................................11

6.7 Storage of isolated ccfDNA .........................................................................................................................................................11

6.7.1 General...................................................................................................................................................................................11

6.7.2 ccfDNA isolated with commercially available kits ...........................................................................12

6.7.3 ccfDNA isolated with the laboratory's own protocols ..................................................................12

Annex A (informative) Impact of pre-examination process steps on circulating cell free DNA

profiles in venous whole blood plasma .....................................................................................................................................13

Bibliography .............................................................................................................................................................................................................................16

© ISO 2019 – All rights reserved iii
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SIST EN ISO 20186-3:2020
ISO 20186-3:2019(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 20186 series 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 2019 – All rights reserved
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SIST EN ISO 20186-3:2020
ISO 20186-3:2019(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 the

pre-examination process, including the specimen collection, transport, storage and processing.

Consequently, this makes the outcome from diagnostics or research unreliable or even impossible

because the subsequent examination might not determine the real situation in the patient, but an

artificial profile generated during the pre-examination processes.

Circulating cell free DNA (ccfDNA) profiles can change significantly after blood collection (e.g. release

of genomic DNA from cells in blood, ccfDNA degradation and fragmentation and ccfDNA quantity

change). Therefore, special measures need to be taken to secure good quality specimens for ccfDNA

[23]

examination. Studies have been undertaken to determine the important influencing factors .

Standardization of the entire workflow from specimen collection to the ccfDNA examination is needed.

This document standardizes the steps of the pre-examination phase of circulating cell free DNA

prepared from plasma of venous whole blood.
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.
© ISO 2019 – All rights reserved v
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SIST EN ISO 20186-3:2020
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SIST EN ISO 20186-3:2020
INTERNATIONAL STANDARD ISO 20186-3:2019(E)
Molecular in vitro diagnostic examinations —
Specifications for pre-examination processes for venous
whole blood —
Part 3:
Isolated circulating cell free DNA from plasma
1 Scope

This document provides recommendations and requirements on the handling, storage, processing

and documentation of venous whole blood specimens intended for circulating cell free DNA (ccfDNA)

examination during the pre-examination phase before an analytical test is performed. This document

covers specimens collected in venous whole blood collection tubes.

This document is applicable to any molecular in vitro diagnostic examination performed by medical

laboratories. It is also intended to be used by laboratory customers, in vitro diagnostics developers and

manufacturers, biobanks, institutions and commercial organizations performing biomedical research,

and regulatory authorities.

Different dedicated measures are taken for stabilizing blood genomic DNA, which are not described in

this document. Blood genomic DNA is covered in ISO 20186-2.

Different dedicated measures are taken for preserving DNA in circulating exosomes, which are not

described in this document.

NOTE ccfDNA obtained from blood by the procedures cited in this document can contain DNA originally

[8][9]
present in exosomes .
DNA in pathogens present in blood is not covered by 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 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
analyte
component represented in the name of a measurable quantity
[SOURCE: ISO 17511:2003, 3.2, modified — The example has been deleted.]
© ISO 2019 – All rights reserved 1
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SIST EN ISO 20186-3:2020
ISO 20186-3:2019(E)
3.2
backflow
flow of a liquid opposite to the usual or desired direction
3.3
blood collection set

intravenous device specialized for venipuncture consisting of a stainless steel beveled needle and tube

(tubing) with attached plastic wings and fitting connector

Note 1 to entry: The connector attaches to an additional blood collection device, e.g. a blood collection tube.

3.4
blood collection tube

tube used for blood collection, usually with a vacuum which forces blood from the vein through the

needle into the tube
3.5
ccfDNA
circulating cell free DNA
extracellular human DNA present in blood and plasma
[8][9]
Note 1 to entry: ccfDNA can include DNA present in vesicles such as exosomes .
3.6
ccfDNA profile
circulating cell free DNA profile

amount of different ccfDNA molecules, present in blood and plasma that can be measured in the absence

of any losses, inhibition and interference
3.7
closed system

non-modifiable system provided by the vendor including all necessary components for the pre-

examination and/or examination (i.e. hardware, software, procedures and reagents)

3.8
DNA
deoxyribonucleic acid

polymer of deoxyribonucleotides occurring in a double-stranded (dsDNA) or single-stranded

(ssDNA) form
[SOURCE: ISO 22174:2005, 3.1.2]
3.9
DNase
deoxyribonuclease
enzyme that catalyzes the degradation of DNA into smaller components
3.10
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 —"analytical test" has been added as additional preferred

term; Notes to entry have been deleted; new Note 1 to entry has been added.]
2 © ISO 2019 – All rights reserved
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SIST EN ISO 20186-3:2020
ISO 20186-3:2019(E)
3.11
examination performance
analytical test performance
analytical performance
ability of an examination procedure to measure or detect a particular analyte

Note 1 to entry: Analytical performance is determined from analytical performance studies used to assess the

ability of an in vitro diagnostic examination procedure to measure or detect a particular analyte.

Note 2 to entry: Analytical performance includes such characteristics as analytical sensitivity, detection limit,

analytical specificity (interference and cross-reactivity), trueness, precision and linearity.

[SOURCE: ISO/TS 17822-1:2014, 3.2, modified — Two preferred terms have been added.]

3.12
examination provider
analytical test provider
entity that provides the specific analytical test
3.13
needle holder

barrel used in routine venipuncture procedures to hold the blood collection tube in place and to protect

the phlebotomist from direct contact with blood
3.14
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 laboratory, isolation of analytes, and end when the analytical

examination begins

Note 1 to entry: The pre-examination phase includes preparative processes, e.g. ccfDNA isolation procedures,

which influence the outcome of the intended examination.

[SOURCE: ISO 15189:2012, 3.15, modified — An additional term has been added and more detail have

been included in the definition; Note 1 to entry has been added.]
3.15
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 have been deleted.]
3.16
proficiency testing

evaluation of participant performance against pre-established criteria by means of inter-laboratory

comparisons
[SOURCE: ISO/IEC 17043:2010, 3.7, modified — Notes have been deleted.]
3.17
RNA
ribonucleic acid

polymer of ribonucleotides occurring in a double-stranded or single-stranded form

[SOURCE: ISO 22174:2005, 3.1.3]
© ISO 2019 – All rights reserved 3
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SIST EN ISO 20186-3:2020
ISO 20186-3:2019(E)
3.18
RNase
ribonuclease
enzyme that catalyzes the degradation of RNA into smaller components
3.19
room temperature
temperature in the range of 18 °C to 25 °C for the purposes of this document
Note 1 to entry: Local or national regulations can have different definitions.
3.20
sample
one or more parts taken from a primary sample
[SOURCE: ISO 15189:2012, 3.24, modified — The example has been deleted.]
3.21
stability

ability of a specimen or sample, when stored under specified conditions, to maintain a stated property

value within specified limits for a specified period of time

[SOURCE: ISO Guide 30:2015, 2.1.15, modified — The words “reference material” have been replaced by

“specimen or sample”.]
3.22
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 term “validated” is used to designate the corresponding status.

[SOURCE: ISO 9000:2015, 3.8.13, modified — Notes 1 and 3 to entry have been deleted, Note 2 to entry

has been renumbered as Note 1 to entry.]
3.23
venous whole blood

blood collected after directly puncturing a vein, usually with a needle and syringe, or other

collection device
3.24
verification

confirmation, through the provision of objective evidence, that specified requirements have been

fulfilled

Note 1 to entry: The term “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;
— reviewing documents prior to issue.

[SOURCE: ISO 9000:2015, 3.8.12, modified — Notes 1 and Note 2 to entry have been deleted; Note 3 to

entry has been renumbered as Note 1 to entry; new Note 2 to entry has been added.]

3.25
workflow
series of activities necessary to complete a task
4 © ISO 2019 – All rights reserved
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SIST EN ISO 20186-3:2020
ISO 20186-3:2019(E)
4 General consideration

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 according to 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 examination result. Thus, the entire workflow,

including specimen/sample storage and transport conditions, and its impact on the stability of

biomolecules intended to be examined shall be verified and validated. Workflow steps which cannot

always be controlled shall be documented and their impact on the examination performance shall

be investigated and mitigation measures shall be established to allow the required examination

performance. In these cases, risk assessment is recommended.

CcfDNA profiles can change significantly after blood collection. The post-collection release of genomic

DNA from cells in blood can change the ccfDNA profile significantly (see A.1). Additional post-

[10][11][12][13]

collection effects can also occur, e.g. ccfDNA fragmentation . All these post-collection

changes can vary individually in specimens from different donors or patients, and they can also

[10][14][15][16]

depend on pathophysiological conditions . This can impact the validity and reliability of the

examination results (see A.2).

Before or during the design of an examination, it shall therefore be investigated and ensured that

the ccfDNA profile(s) intended to be analysed is/are not compromised in a manner impacting the

examination performance. This can be done, e.g. by applying the intended examination to specimens/

samples which underwent time course studies reflecting the individual pre-examination process steps

such as transport and storage and by implementing measures to prevent or reduce impacts by the

identified pre-analytical variables, e.g. by using blood collection tubes with stabilizers.

Safety procedures for handling and transport shall be in place. Safety requirements on transport and

handling shall be considered (see ISO 15189 and ISO 15190).

During the whole pre-examination process, precautions shall be taken to avoid cross contamination

between different samples/specimens, 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 validation, verification, use and performance lies with the user.
5 Outside the laboratory
5.1 Specimen collection
5.1.1  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 or patient [e.g. healthy, disease type, concomitant

disease, demographics (e.g. age and gender)];

NOTE In particular, e.g. cancer, inflammation, diabetes, hepatic disease, coronary disease, respiratory

[10]

syndrome, trauma, after exhaustive exercise , in elderly patients suffering from acute or chronic disease,

first trimester of pregnancy, placental disorders as pre-term labour, pre-eclampsia and malimplantation

[10][14][15][16]
have been reported to affect both ccfDNA quantity and fragmentation .

b) the information about medical treatment and special treatment prior to blood collection (e.g.

anaesthetics, medications, fasting status);
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SIST EN ISO 20186-3:2020
ISO 20186-3:2019(E)
c) the type and purpose of the proposed examination requested;
d) the appropriate consent from the specimen donor/patient.
See also ISO 15189:2012, 5.4.4.
5.1.2  Selection of the venous whole blood collection tube by the laboratory

The ccfDNA profile can be influenced by inadequate venous whole blood collection procedures and

...

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