CEN/TS 16835-2:2015
(Main)Molecular in vitro diagnostic examinations - Specifications for pre-examination processes for venous whole blood - Part 2: Isolated genomic DNA
Molecular in vitro diagnostic examinations - Specifications for pre-examination processes for venous whole blood - Part 2: Isolated genomic DNA
This Technical Specification recommends the handling, documentation and processing of venous whole blood specimens intended for genomic DNA analysis during the preanalytical phase before a molecular assay is performed. This Technical Specification covers specimens collected by venous whole blood collection tubes. This Technical Specification is applicable to molecular in vitro diagnostic examinations (e.g. in vitro diagnostic laboratories, laboratory customers, in vitro diagnostics developers and manufacturers, institutions and commercial organizations performing biomedical research, biobanks, and regulatory authorities).
Blood genomic DNA can fragment or degrade after blood collection. Therefore, special measures need to be taken to secure good quality blood samples for genomic DNA analysis. This is particularly relevant for analytical test procedures requiring high molecular weight DNA.
Different dedicated measures need to be taken for preserving blood cell free circulating DNA, which are not described in this Technical Specification. Circulating cell free DNA in blood is covered in CEN/TS 16835-3, Molecular in vitro diagnostic examinations -Specifications for pre-examination processes for venous whole blood - Part 3: Isolated circulating cell free DNA from plasma.
Different dedicated measures need to be taken for collecting, stabilizing, transporting and storing capillary blood as well as for blood collected and stored by paper based technologies. These are not described in this Technical Specification.
Pathogen DNA present in blood is not covered by this Technical Specification.
Molekularanalytische in-vitro-diagnostische Verfahren - Spezifikationen für präanalytische Prozesse für venöse Vollblutproben - Teil 2: Isolierte genomische DNS
Diese Technische Spezifikation gibt Empfehlungen zur Handhabung, Dokumentation und Verarbeitung von für die Analyse genomischer DNS vorgesehenen venösen Vollblutproben während der präanalytischen Phase vor Beginn der molekularen Analyse. Diese Technische Spezifikation behandelt Untersuchungsmaterial, das mit Blutentnahmeröhrchen für venöses Vollblut entnommen wurde. Diese Technische Spezifikation gilt für molekulare in vitro diagnostische Untersuchungen (z. B. In vitro Diagnostik Labore, Kunden dieser Labore, Entwickler und Hersteller von In vitro Diagnostika, Einrichtungen und kommerzielle Organisationen, die in der biomedizinischen Forschung tätig sind, Biobanken und Aufsichtsbehörden).
Bei genomischer DNS aus dem Blut kann es nach der Blutentnahme zu einer Fragmentierung oder einem Abbau kommen. Daher müssen besondere Maßnahmen getroffen werden, um eine gute Qualität der Blutproben für die Analyse der genomischen DNS sicherzustellen. Dies trifft besonders bei analytischen Prüfverfahren zu, die DNS mit einer hohen Molekülmasse erfordern.
Zur Konservierung zellfreier zirkulierender DNS in Blut müssen gesonderte Maßnahmen getroffen werden, die nicht in dieser Technischen Spezifikation beschrieben sind. Zirkulierende zellfreie DNS in Blut ist behandelt in CEN/TS 16835-3, Molekularanalytische in vitro diagnostische Verfahren - Spezifikation für präanalytische Prozesse für venöse Vollblutproben - Aus Plasma isolierte zirkulierende zellfreie DNS.
Nicht durch diese Technische Spezifikation abgedeckt ist in Blut vorhandene pathogene DNS.
Tests de diagnostic moléculaire in vitro - Spécifications relatives aux processus pré-analytiques pour le sang total veineux - Partie 2: ADN génomique extrait
La présente Spécification technique donne des recommandations pour la manipulation, la documentation et le traitement des prélèvements de sang total veineux destinés à l’analyse de l’ADN génomique pendant la phase pré-analytique, avant d’effectuer un essai moléculaire. Elle concerne les spécimens collectés dans des tubes de prélèvement de sang total veineux. La présente Spécification technique est applicable aux tests de diagnostic moléculaire in vitro (par exemple laboratoires de diagnostic in vitro, clients de laboratoires, concepteurs et fabricants de tests de diagnostics in vitro, institutions et organisations commerciales travaillant dans la recherche biomédicale, les biobanques et les autorités réglementaires).
L’ADN génomique sanguin peut se fragmenter ou se dégrader après le prélèvement sanguin. Par conséquent, des mesures spécifiques doivent être prises pour assurer la bonne qualité des échantillons de sang en vue de l’analyse de l’ADN génomique, en particulier pour les procédures d’essai analytique nécessitant un ADN de haut poids moléculaire.
Des mesures spécifiques, non décrites dans la présente Spécification technique, doivent être prises pour préserver l’ADN libre circulant dans le sang. L’ADN libre circulant dans le sang est traité dans la FprCEN/TS 16835-3, Tests de diagnostic moléculaire in vitro -Spécifications relatives aux processus pré-analytiques pour le sang total veineux - Partie 3 : ADN libre circulant extrait du plasma.
Des mesures spécifiques doivent être prises pour prélever, stabiliser, transporter et stocker le sang capillaire, et pour prélever et stocker le sang par des technologies sur support papier. Ces mesures ne sont pas décrites dans la présente Spécification technique.
L’ADN pathogène présent dans le sang n’est pas couvert par la présente Spécification technique.
Molekularne diagnostične preiskave in vitro - Specifikacije za predpreiskovalne procese za vensko polno kri - 2. del: DNA, izoliran iz genoma
Ta tehnična specifikacija vsebuje priporočila za obravnavo, dokumentiranje in obdelavo vzorcev venske polne krvi, namenjenih za analizo genomskega DNA-ja med predanalizno fazo, preden se izvede molekularni preskus. Ta tehnična specifikacija zajema vzorce, ki so zbrani s cevkami za zbiranje venske polne krvi. Ta tehnična specifikacija se uporablja za molekularne diagnostične preiskave in vitro (npr. diagnostični laboratoriji in vitro, laboratorijske stranke, razvijalci in proizvajalci diagnostike in vitro, institucije in komercialne organizacije, ki izvajajo biomedicinske raziskave, biobanke ter regulativni organi).
Krvni genomski DNA lahko po odvzemu krvi razpade ali se razkroji. Zato je treba uporabiti posebne ukrepe za pridobivanje vzorcev krvi dobre kakovosti za analizo genomskega DNA-ja. To je še posebej pomembno za analitične preskusne postopke, ki zahtevajo DNA z veliko molekularno težo.
Za ohranjanje cirkulirajočega brezceličnega DNA-ja je treba uporabiti drugačne namenske ukrepe, ki niso opisani v tem tehničnem poročilu. Cirkulirajoči brezcelični DNA je opisan v standardu CEN/TS 16835-3, Molekularne diagnostične preiskave in vitro - Specifikacije za predpreiskovalne procese za vensko polno kri - 3. del: Iz plazme izolirani cirkulirajoči brezcelični DNA
Za zbiranje, stabiliziranje, prevoz in shrambo kapilarne krvi in krvi, ki se zbira in shranjuje s tehnologijami, ki temeljijo na papirju, je treba uporabiti drugačne namenske ukrepe. Ti niso opisani v tej tehnični specifikaciji.
Patogeni DNA v krvi ni zajet v tej tehnični specifikaciji.
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
01-december-2015
0ROHNXODUQHGLDJQRVWLþQHSUHLVNDYHLQYLWUR6SHFLILNDFLMH]DSUHGSUHLVNRYDOQH
SURFHVH]DYHQVNRSROQRNULGHO '1$L]ROLUDQL]JHQRPD
Molecular in vitro diagnostic examinations - Specifications for pre-examination processes
for venous whole blood - Part 2: Isolated genomic DNA
Molekularanalytische in-vitro-diagnostische Verfahren - Spezifikationen für
präanalytische Prozesse für venöse Vollblutproben - Teil 2: Isolierte genomische DNA
Ta slovenski standard je istoveten z: CEN/TS 16835-2:2015
ICS:
11.100.10 'LDJQRVWLþQLSUHVNXVQL In vitro diagnostic test
VLVWHPLLQYLWUR systems
11.100.30 Analiza krvi in urina Analysis of blood and urine
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
CEN/TS 16835-2
TECHNICAL SPECIFICATION
SPÉCIFICATION TECHNIQUE
October 2015
TECHNISCHE SPEZIFIKATION
ICS 11.100.30
English Version
Molecular in vitro diagnostic examinations - Specifications
for pre-examination processes for venous whole blood -
Part 2: Isolated genomic DNA
Tests de diagnostic moléculaire in vitro - Spécifications Molekularanalytische in-vitro-diagnostische Verfahren
relatives aux processus pré-analytiques pour le sang - Spezifikationen für präanalytische Prozesse für
total veineux - Partie 2: ADN génomique extrait venöse Vollblutproben - Teil 2: Isolierte genomische
DNS
This Technical Specification (CEN/TS) was approved by CEN on 31 August 2015 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, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, 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: Avenue Marnix 17, B-1000 Brussels
© 2015 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN/TS 16835-2:2015 E
worldwide for CEN national Members.
Contents Page
European foreword . 3
Introduction . 4
1 Scope . 5
2 Normative references . 5
3 Terms and definitions . 5
4 General considerations . 7
5 Outside the laboratory . 7
5.1 Primary venous whole blood collection manual . 7
5.1.1 Information about the primary sample donor . 7
5.1.2 Selection of the venous whole blood collection tube by the laboratory . 8
5.1.3 Primary venous whole blood sample collection from the patient and stabilization
procedures . 8
5.1.4 Information on the primary blood sample and storage requirements at the blood
collection facility . 8
5.2 Transport requirements. 9
6 Inside the laboratory . 10
6.1 Primary sample reception . 10
6.2 Storage requirements . 10
6.3 Isolation of the genomic DNA . 11
6.3.1 General . 11
6.3.2 Using commercial kits . 12
6.3.3 Using the laboratories own protocols . 12
6.4 Quantity and quality assessment of isolated genomic DNA . 12
6.5 Storage of isolated genomic DNA . 13
Annex A (informative) Impact of preanalytical workflow steps on venous whole blood
genomic DNA quality . 14
A.1 General information on operated experiments in Annex A . 14
A.2 Influence of preanalytical variables (blood storage duration and temperature, and
DNA isolation methods) on genomic DNA integrity . 14
A.3 Influence of blood storage time on the genomic DNA integrity. 15
A.4 Influence of genomic DNA integrity on an analytical test based on long PCR
amplicons . 17
A.5 Influence of blood storage conditions on the performance of PCR tests based on
short amplicons . 18
Bibliography . 20
European foreword
This document (CEN/TS 16835-2:2015) 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 [and/or CENELEC] shall not be held responsible for identifying any or all such patent
rights.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the
following countries are bound to announce this Technical Specification: 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, 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 signatures of nucleic acids, proteins, and metabolites in human
tissues and body fluids. However, the profiles of these molecules can change drastically during primary
sample 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.
A standardization of the entire process from primary sample collection to genomic DNA analysis is
needed due to genomic DNA degradation and fragmentation after blood collection. Studies have been
undertaken to determine the important influencing factors. This Technical Specification draws upon
such work to codify and standardize the steps for venous whole blood genomic DNA analysis in what is
referred to as the preanalytical phase.
1 Scope
This Technical Specification recommends the handling, documentation and processing of venous whole
blood specimens intended for genomic DNA analysis during the preanalytical phase before a molecular
assay is performed. This Technical Specification covers specimens collected by venous whole blood
collection tubes. This Technical Specification is applicable to molecular in vitro diagnostic examinations
(e.g. in vitro diagnostic laboratories, laboratory customers, in vitro diagnostics developers and
manufacturers, institutions and commercial organizations performing biomedical research, biobanks,
and regulatory authorities).
Blood genomic DNA can fragment or degrade after blood collection. Therefore, special measures need to
be taken to secure good quality blood samples for genomic DNA analysis. This is particularly relevant
for analytical test procedures requiring high molecular weight DNA.
Different dedicated measures need to be taken for preserving blood circulating cell free DNA, which are
not described in this Technical Specification. Circulating cell free DNA in blood is covered in
CEN/TS 16835-3, Molecular in vitro diagnostic examinations — Specifications for pre-examination
processes for venous whole blood — Part 3: Isolated circulating cell free DNA from plasma.
Different dedicated measures need to be taken for collecting, stabilizing, transporting and storing
capillary blood as well as for blood collected and stored by paper based technologies. These are not
described in this Technical Specification.
DNA from pathogens present in blood is not covered by this Technical Specification.
2 Normative references
The following documents, in whole or in part, are normatively referenced in this document and are
indispensable for its application. For dated references, only the edition cited applies. For undated
references, the latest edition of the referenced document (including any amendments) applies.
EN ISO 15189:2012, Medical laboratories — Requirements for quality and competence (ISO 15189:2012,
Corrected version 2014-08-15)
ISO 15190, Medical laboratories — Requirements for safety
3 Terms and definitions
For the purposes of this document, the terms and definitions given in EN ISO 15189:2012 and the
following apply.
3.1
ambient temperature
unregulated temperature of the surrounding air
3.2
analytical phase
processes that start with the isolated analyte and include all kind of parameter testing or chemical
manipulation for quantitative or qualitative analysis
3.3
blood genomic DNA stabilizers
compounds, solutions or mixtures that are made to minimize degradation and fragmentation of
genomic DNA in a blood sample
3.4
DNA
deoxyribonucleic acid
polymer of deoxyribonucleotides occurring in a double-stranded (dsDNA) or single-stranded (ssDNA)
form
[SOURCE: EN ISO 22174:2005, 3.1.2]
3.5
genomic DNA
DNA from the genome containing all coding (exon) and non-coding (intron and other) sequences
Note 1 to entry: In this document it is always only referred to genomic DNA present in blood cells, excluding
circulating cell free DNA.
3.6
high molecular weight DNA
HMW DNA
DNA larger than 50 kb for the purpose of this document
3.7
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), temporary
storage, transportation to and within the analytical laboratory, aliquotting, retrieval, isolation of
analytes, and end when the analytical examination begins
[SOURCE: EN ISO 15189:2012, 3.15, modified — An additional term was added and more details were
included.]
Note 1 to entry: The preanalytical phase may include preparative processes that may influence the outcome of
the intended examination.
3.8
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: EN ISO 15189:2012, 3.16, modified — The term and definition is used here without the
original notes.]
3.9
room temperature
temperature which is defined as 18 °C to 25 °C for the purpose of
...
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