CEN/TS 17747:2022
(Main)Molecular in vitro diagnostic examinations - Specifications for pre-examination processes for exosomes and other extracellular vesicles in venous whole blood - DNA, RNA and proteins
Molecular in vitro diagnostic examinations - Specifications for pre-examination processes for exosomes and other extracellular vesicles in venous whole blood - DNA, RNA and proteins
This document gives guidelines on the handling, storage, processing and documentation of venous whole blood specimens intended for DNA, RNA and protein examination from exosomes and other extracellular vesicles during the pre-examination phase before a molecular examination is performed. This document covers specimens collected in venous whole blood collection tubes.
The pre-examination process described in this document results in isolated DNA, RNA and proteins from enriched exosomes and other extracellular vesicles.
This document is applicable to molecular in vitro diagnostic examinations performed by medical laboratories. It is also intended to be used by health care institutions including facilities collecting and handling specimen, laboratory customers, in vitro diagnostics developers and manufacturers, biobanks, institutions and commercial organizations performing biomedical research, and regulatory authorities.
Different dedicated measures are taken during the pre-examination phase for venous whole blood circulating cell-free RNA (ccfRNA) examination and for venous whole blood circulating cell-free DNA (ccfDNA) examination, both without prior enrichment of exosomes and other extracellular vesicles. These are not described in this document but are covered in EN ISO 20186 3, Molecular in vitro diagnostic examinations - Specifications for pre-examination processes for venous whole blood - Part 3: Isolated circulating cell free DNA from plasma and CEN/TS 17742, Molecular in vitro diagnostic examinations - Specifications for pre-examination processes for venous whole blood - Isolated circulating cell free RNA from plasma.
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 Exosomen und andere extrazelluläre Vesikel im venösen Vollblut - DNA, RNA und Proteine
Dieses Dokument gibt Empfehlungen zur Handhabung, Lagerung, Verarbeitung und Dokumentation von Proben venösen Vollbluts, die für die Untersuchung von DNA, RNA und Proteinen aus Exosomen und anderen extrazellulären Vesikeln während der präanalytischen Phase vor der Durchführung einer molekularen Analyse vorgesehen sind. Dieses Dokument betrifft Untersuchungsmaterial, das mit Blutentnahmeröhrchen für venöses Vollblut entnommen wurde.
Mit dem in diesem Dokument beschriebenen präanalytischen Verfahren werden DNA, RNA und Proteine aus angereicherten Exosomen und anderen extrazellulären Vesikeln isoliert.
Dieses Dokument ist anwendbar auf molekulare in vitro-diagnostische Untersuchungen, die in medizinischen Laboratorien durchgeführt werden. Es ist darüber hinaus für die Verwendung durch Gesundheits¬einrichtungen, einschließlich Einrichtungen, die Untersuchungsmaterial entnehmen und handhaben, Laborkunden, Entwickler und Hersteller von In vitro-Diagnostika, Biobanken, in der biomedizinischen Forschung tätige Einrichtungen und kommerzielle Organisationen sowie Aufsichtsbehörden, vorgesehen.
Für die während der präanalytischen Phase durchgeführte Untersuchung von zirkulierender zellfreier RNA (ccfRNA, en: circulating cell-free RNA) und zirkulierender zellfreier DNA (ccfDNA, en: circulating cell-free DNA) aus venösem Vollblut, jeweils ohne vorherige Anreicherung von Exosomen und anderen extrazellulären Vesikeln, werden spezielle andere Maßnahmen angewendet. Diese werden nicht in diesem Dokument beschrieben, sind jedoch Gegenstand von EN ISO 20186 3, Molekularanalytische in-vitro-diagnostische Ver¬fahren — Spezifikationen für präanalytische Prozesse für venöse Vollblutproben — Teil 3: Aus Plasma isolierte zirkulierende zellfreie DNA, und CEN/TS 17742, Molekularanalytische in-vitro-diagnostische Verfahren — Spezifikationen für präanalytische Prozesse für venöse Vollblutproben — Isolierte zirkulierende zellfreie RNA aus Plasma.
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 processuses préanalytiques pour exosomes et autres vésicules extracellulaires dans le sang total veineux - ADN, ARN et protéines
Molekularne diagnostične preiskave in vitro - Specifikacije za predpreiskovalne procese za eksosome in druge zunajcelične vezikle v vensko polni krvi - DNK, RNK in proteini
Ta dokument podaja smernice glede obravnave, shranjevanja, obdelave in dokumentiranja vzorcev venske polne krvi, namenjenih za analizo DNK, RNK in beljakovin iz eksosomov in drugih zunajceličnih veziklov v predpreiskovalnem procesu, preden se izvede molekularna preiskava. Ta dokument zajema vzorce, zbrane v epruvete za odvzem polne venske krvi.
V predpreiskovalnem procesu, opisanem v tem dokumentu, se pridobijo izolirana DNK, RNK in beljakovine iz obogatenih eksosomov ter drugih zunajceličnih veziklov.
Ta dokument se uporablja za molekularne diagnostične preiskave in vitro, ki jih izvajajo v medicinskih laboratorijih. Namenjen je tudi temu, da ga uporabljajo laboratorijske stranke, razvijalci in proizvajalci diagnostike in vitro, biobanke, institucije in komercialne organizacije, ki izvajajo biomedicinske raziskave, ter regulativni organi.
Med predpreiskovalnim procesom se izvajajo različni namenski ukrepi za preiskavo cirkulirajoče brezcelične RNK (ccfRNA) polne krvi in za preiskavo cirkulirajoče brezcelične DNK (ccfDNA) venske krvi, pri obeh brez predhodne obogatitve eksosomov in drugih zunajceličnih veziklov. Ti niso opisani v tem dokumentu, ampak jih vsebuje standard prEN ISO 20186-3, Molekularne diagnostične preiskave in vitro – Specifikacije za predpreiskovalne procese za vensko polno kri – 3. del: Iz plazme izolirana cirkulirajoča brezcelična RNK in CEN/PWI, Molekularne diagnostične preiskave in vitro – Specifikacije za predpreiskovalne procese za vensko polno kri – Iz plazme izolirana cirkulirajoča brezcelična RNK.
OPOMBA: Za določene teme, ki so zajete v tem dokumentu, lahko veljajo tudi mednarodni, nacionalni ali regionalni predpisi ali zahteve.
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-junij-2022
Molekularne diagnostične preiskave in vitro - Specifikacije za predpreiskovalne
procese za eksosome in druge zunajcelične vezikle v vensko polni krvi - DNK, RNK
in proteini
Molecular in vitro diagnostic examinations - Specifications for pre-examination processes
for exosomes and other extracellular vesicles in venous whole blood - DNA, RNA and
proteins
Molekularanalytische in-vitro-diagnostische Verfahren - Spezifikationen für
präanalytische Prozesse für Exosomen und andere extrazelluläre Vesikel im venösen
Vollblut - DNA, RNA und Proteine
Analyses de diagnostic moléculaire in vitro - Spécifications relatives aux processuses
préanalytiques pour exosomes et autres vésicules extracellulaires dans le sang total
veineux - ADN, ARN et protéines
Ta slovenski standard je istoveten z: CEN/TS 17747:2022
ICS:
11.100.10 Diagnostični preskusni In vitro diagnostic test
sistemi in vitro systems
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
CEN/TS 17747
TECHNICAL SPECIFICATION
SPÉCIFICATION TECHNIQUE
April 2022
TECHNISCHE SPEZIFIKATION
ICS 11.100.10
English Version
Molecular in vitro diagnostic examinations - Specifications
for pre-examination processes for exosomes and other
extracellular vesicles in venous whole blood - DNA, RNA
and proteins
Analyses de diagnostic moléculaire in vitro - Molekularanalytische in-vitro-diagnostische Verfahren
Spécifications relatives aux processuses préanalytiques - Spezifikationen für präanalytische Prozesse für
pour exosomes et autres vésicules extracellulaires Exosomen und andere extrazelluläre Vesikel im
dans le sang total veineux - ADN, ARN et protéines venösen Vollblut - DNA, RNA und Proteine
This Technical Specification (CEN/TS) was approved by CEN on 13 March 2022 for provisional application.
The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to
submit their comments, particularly on the question whether the CEN/TS can be converted into a European Standard.
CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS
available promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in
parallel to the CEN/TS) until the final decision about the possible conversion of the CEN/TS into an EN is reached.
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
© 2022 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN/TS 17747:2022 E
worldwide for CEN national Members.
Contents Page
European foreword . 4
Introduction . 4
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 7
4 General Requirements . 11
5 Outside the laboratory . 13
5.1 Specimen collection . 13
5.1.1 General . 13
5.1.2 Information about the patient/specimen donor . 13
5.1.3 Selection of the venous whole blood collection tube by the laboratory . 13
5.1.4 Venous whole blood specimen collection from the patient/donor . 14
5.2 Specimen storage and transport . 15
5.2.1 General . 15
5.2.2 Storage and transport using blood collection tubes with stabilizers . 15
5.2.3 Storage and transport using blood collection tubes without stabilizers . 15
6 Inside the laboratory . 16
6.1 Specimen reception . 16
6.2 Specimen storage after transport and reception . 16
6.3 Plasma preparation . 16
6.4 Storage requirements for plasma samples . 17
6.5 Enrichment of EVs from specimen . 17
6.5.1 General . 17
6.5.2 Using a commercial EV enrichment system . 18
6.5.3 Using the laboratory’s own EV enrichment procedure . 18
6.5.4 Quality of enriched EVs . 18
6.5.5 Storage of enriched EVs . 19
6.6 Isolation of analyte of interest from EVs . 19
6.6.1 General . 19
6.6.2 Using a commercial kit for EV DNA, RNA, protein isolation intended for diagnostic
use . 19
6.6.3 Using a laboratory developed isolation procedure . 20
6.7 Quantity and quality assessment of isolated analyte of interest . 21
6.7.1 General . 21
6.7.2 Quantity assessment of EV DNA and RNA . 21
6.7.3 Quality assessment of DNA and RNA . 22
6.7.4 Quantity and quality assessment of EV proteins . 22
6.8 Storage of isolated analyte of interest . 23
6.8.1 General . 23
6.8.2 Storage of DNA . 23
6.8.3 Storage of RNA . 24
6.8.4 Storage of EV protein . 25
Annex A (informative) Example of a typical protocol for plasma preparation for EV
enrichment from unstabilized blood . 26
Annex B (informative) Overview of different enrichment procedures . 27
B.1 General . 27
B.2 EV enrichment by centrifugation . 27
B.3 EV enrichment by filtration . 27
B.4 EV enrichment by chromatography . 28
B.5 EV enrichment by precipitation . 28
B.6 EV enrichment by immunoaffinity . 28
B.7 EV enrichment by microfluidics . 28
Bibliography . 29
European foreword
This document (CEN/TS 17747:2022) has been prepared by Technical Committee CEN/TC 140 “In vitro
diagnostic medical devices”, the secretariat of which is held by DIN.
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.
Any feedback and questions on this document should be directed to the users’ national standards body.
A complete listing of these bodies can be found on the CEN website.
According to the CEN/CENELEC Internal Regulations, the national standards organisations of the
following countries are bound to announce this Technical Specification: 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.
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 process.
Besides cell free circulating nucleic acids, circulating tumour cells (CTCs) and other rare cells, exosomes
and other extracellular vesicles represent another key component of liquid biopsies. Therefore, there is
a strongly increasing interest in research and emerging diagnostics in exosomes and other extracellular
vesicles.
The pre-examination process described in this document results in enriched extracellular vesicles (EV)
(e.g. exosomes) or DNA, RNA and proteins isolated therefrom.
New additional extracellular vesicles can be released and existing extracellular vesicles can be lost after
blood collection, thus changing the overall EV DNA/RNA/protein profiles. Also, different anticoagulants
in different types of blood collection tubes can influence the release of EVs from different cells present in
blood, including those from platelets. Further factors can influence the post collection changes of the
entire blood EV composition, such as storage and transport temperature and duration, centrifugation
parameters, etc.
Standardization of the entire workflow from the specimen collection to the EV surface protein and
isolated DNA, RNA and protein examination from EVs is therefore needed. Studies have been undertaken
to determine the important influencing factors. This document draws upon such work to codify and
standardize the steps of EV surface pr
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