SIST-TS 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 DNA
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
SIST-TS CEN/TS 16835-2:2015
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
SIST-TS CEN/TS 16835-2:2015 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
---------------------- Page: 1 ----------------------
SIST-TS CEN/TS 16835-2:2015
---------------------- Page: 2 ----------------------
SIST-TS CEN/TS 16835-2:2015
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.
---------------------- Page: 3 ----------------------
SIST-TS CEN/TS 16835-2:2015
CEN/TS 16835-2:2015 (E)
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
2
---------------------- Page: 4 ----------------------
SIST-TS CEN/TS 16835-2:2015
CEN/TS 16835-2:2015 (E)
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.
3
---------------------- Page: 5 ----------------------
SIST-TS CEN/TS 16835-2:2015
CEN/TS 16835-2:2015 (E)
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.
4
---------------------- Page: 6 ----------------------
SIST-TS CEN/TS 16835-2:2015
CEN/TS 16835-2:2015 (E)
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
5
---------------------- Page: 7 ----------------------
SIST-TS CEN/TS 16835-2:2015
CEN/TS 16835-2:2015 (E)
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 this document
6
---------------------- Page: 8 ----------------------
SIST-TS CEN/TS 16835-2:2015
CEN/TS 16835-2:2015 (E)
3.10
stability
ability of a sample material, 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” were replaced by
“sample material".]
Note 1 to entry: The measured constituent for the purpose of this document is genomic DNA.
4 General considerations
For general statements on primary sample collection and handling (including avoidance of cross
contaminations), see EN ISO 15189:2012, 5.2.6, 5.4.4. Consumables including kits shall be verified
before use in examination (see EN ISO 15189:2012, 5.3.2.3); EN ISO 15189:2012, 5.5.1.2 and 5.5.1.3 can
also apply.
As all steps of a diagnostic workflow can influence the final analytical performance, the entire workflow,
comprising the preanalytical steps, including information on sample stability and storage conditions,
and the analytical steps should be verified and validated (see EN ISO 15189).
The stability of the genomic DNA should be investigated throughout the complete pre-analytical
workflow.
Before or during the design of the analytical test system it should be investigated and ensured that the
genomic DNA minimum amount and size required for the analytical test are not affected by the
envisioned entire preanalytical workflow.
If a commercial product is not used in accordance with the manufacturers' instructions, responsibility
for its use and performance lies with the user.
Safety regulations on facilities, transport and handling shall be considered (EN ISO 15189:2012, 5.2.3
and 5.4.5, and ISO 15190).
5 Outside the laboratory
5.1 Primary venous whole blood collection manual
5.1.1 Information about the primary sample donor
The documentation should include, but is not limited to:
a) the primary donor / patient ID, which can be in the form of a code;
b) the health status and relevant lifestyle factors of the blood donor (e.g. healthy, disease type, gender,
age);
c) the information about medical treatment and special treatment prior to blood collection (e.g.
anaesthetics, medications);
d) the type and the purpose of the analytical test requested.
See also EN ISO 15189:2012, 5.4.4.
7
---------------------- Page: 9 ----------------------
SIST-TS CEN/TS 16835-2:2015
CEN/TS 16835-2:2015 (E)
5.1.2 Selection of the venous whole blood collection tube by the laboratory
The quality of genomic DNA can be influenced (e.g., DNA fragmentation), by inadequate blood collection
procedures, inappropriate storage/shipping conditions and DNA isolation procedures, [3], [4], [5], [6],
[7], [8], [9], [10].
Blood should be collected in appropriate venous whole blood collection tubes containing an
anticoagulant such as EDTA or Acid Citrate Dextrose (ACD) [11].
NOTE Blood collection tubes containing EDTA as an anticoagulant are preferable for most genomic DNA
analysis. Blood collection tubes containing heparin as an anticoagulant can impact the purity of the isolated
genomic DNA, when using genomic DNA isolation methods not eliminating the heparin. Carrying over of heparin
into the genomic DNA eluate can cause inhibitions in analytical test technologies, such as PCR.
Specifically developed blood collection tubes, containing genomic DNA stabilizing reagents, are also
available aimed to standardize blood collection, transport and storage of venous whole blood.
5.1.3 Primary venous whole blood sample collection from the patient and stabilization
procedures
1. The identity of the person collecting the primary sample and the time of blood collection according
to EN ISO 15189:2012, 5.4.4.3, f) shall be documented.
2. For the labelling (sample identification) of the blood collection tube a routine procedure
(EN ISO 15189:2012, 5.4.4.3, e)) or a procedure with additional information (e.g. 2D-barcode) shall
be used.
3. Standard venepuncture technique can be used. Steps for preventing possible backflow may be
required. The manufacturers’ instructions for using the blood collection tubes shall be followed. A
blood collection set and needle holder can be required when using blood genomic DNA stabilizer
containing tubes. In this case, the instructions of the collection set and needle holder manufacturer
shall be followed.
NOTE There is no known specific effect of venous whole blood draw procedure on the genomic DNA.
Routine procedures can therefore be used.
4. Blood collection tubes shall be filled in accordance to the manufacturers’ instructions and attention
should be drawn to the correct positioning of the collection tube during the blood draw as well as
the required volume.
5. Blood collection tube manufacturers' instructions for mixing or inverting the tube immediately
after blood collection shall be followed.
NOTE Unless additives are homogenously mixed with the blood sample, the genomic DNA quality can be
compromised, which can impact the validity and reliability of the analytical test results.
6. Any tampering with and/or additions to the primary sample shall be documented.
5.1.4 Information on the primary blood sample and storage requirements at the blood
collection facility
5.1.4.1 General
As blood genomic DNA can fragment or degrade after blood collection (Figure A.1) and can thereby
affect the validity and reliability of the analytical test result (Figure A.3), the documentation on the
primary blood sample shall include the date and should also include the time of blood collection [11].
8
---------------------- Page: 10 ----------------------
SIST-TS CEN/TS 16835-2:2015
CEN/TS 16835-2:2015 (E)
For samples dedicated to be archived in a biobank it is usually not known which individual genomic
DNA tests will be performed after archiving, therefore either tubes with genomic DNA stabilizers should
be used or, if using tubes without genomic DNA stabilizers, the recommendations for HMW DNA should
be followed (see Table 1 and 5.1.4.3.2).
The temporary storage duration in the blood collection facility contributes to the total duration for
storage.
5.1.4.2 Using blood collection tubes with stabilizers
For storing the primary blood samples collected in blood collection tubes with blood genomic DNA
stabilizers, the blood collection tube manufacturers' instructions on storage conditions shall be
followed (including the storage duration and temperature). Where the analytical test providers'
instructions are more stringent, these shall be followed. The storage conditions (including the storage
duration and temperature) shall be documented.
5.1.4.3 Using blood collection tubes without stabilizers
5.1.4.3.1 When using blood collection tubes without blood genomic DNA stabilizers, the analytical
test providers' instructions on storage conditions shall be followed. This can require documentation of
storage conditions.
5.1.4.3.2 When using blood collection tubes without blood genomic DNA stabilizers and no
requirements on the storage conditions are available from the analytical test provider, the primary
venous whole blood samples should be processed as soon as possible.
As blood collection tubes containing EDTA as an anticoagulant are most wide spread for genomic DNA
analysis the following recommendations (see also Table 1) refer to this blood collection tube type. For
analytical tests requiring High Molecular Weight DNA, the blood sample should be stored at room
temperature for not longer than one day or at 2 °C to 8 °C for not longer than three days. For a longer
storage the sample should be kept at −20 °C for not longer than 1 month, or at −70 °C or below for
longer storage. For the analysis of DNA variants not requiring HMW DNA analyses, the blood sample
should be stored at room temperature for up to 3 days or at 2 °C to 8 °C for up to 7 days (Figure A.4).
For a longer storage the sample should be kept at −20 °C for up to 3 months or at −70 °C or below for
longer storage.
The storage conditions (including the storage duration and temperature) shall be documented.
5.2 Transport requirements
The required transport conditions shall be documented including any deviations therefrom.
When using blood collection tubes with blood genomic DNA stabilizers, the tubes' manufacturers'
instructions on transport conditions shall be followed (including the transport duration and
temperature). Where the analytical test providers' instructions are more stringent, these shall be
followed.
When using blood collection tubes without genomic DNA stabilizers, the analytical test providers´
instructions on transport conditions shall be followed. This can require the documentation of transport
conditions (including the transport duration and temperature).
When using blood collection tubes without genomic DNA stabilizers and no analytical test provider's
instructions are available, the primary blood sample should be transported at either room temperature,
at 2 °C to 8 °C, or at −20 °C or below within the specifications given in 5.1.4.3.2 and Table 1 in order to
minimize the degradation and fragmentation of the blood genomic DNA [11], [12].
See also EN ISO 15189:2012, 5.4.5.
The transport duration to the laboratory contributes to the total duration for storage.
9
---------------------- Page: 11 ----------------------
SIST-TS CEN/TS 16835-2:2015
CEN/TS 16835-2:2015 (E)
6 Inside the laboratory
6.1 Primary sample reception
The blood sample reception time shall be documented. Nonconformities of labelling, transport
conditions and blood volume differences to specifications, leaking/broken tubes etc. shall be
documented.
NOTE This includes for example a note, when samples shipped on wet-ice were not cool anymore or
transportation containers do not contain dry-ice as intended by the sender.
Where there are nonconformities in labelling, transport conditions, overall storage and transport
duration or blood volume that could affect the validity and reliability of the analytical test result, a new
sample should be obtained.
6.2 Storage requirements
The storage temperature and time interval between primary sample receipt and sample processing for
genomic DNA isolation shall be documented. Storage temperature and total storage duration shall not
exceed specifications identified in 5.1.4.2, 5.1.4.3.1 and 5.2, and should not exceed specifications
identified in 5.1.4.3.2.
The primary blood sample total storage duration shall include the duration for storage at the blood
collection facility (5.1.4), for transportation to the laboratory (5.2) and for further storage at the
laboratory or other institutions. Any specified maximum storage duration given by the blood collection
tube manufacturer or the analytical test manufacturer shall not be exceeded. If such specifications are
not available, the maximum storage duration shall be validated and generally kept to a minimum.
See also Table 1.
10
---------------------- Page: 12 ----------------------
SIST-TS CEN/TS 16835-2:2015
CEN/TS 16835-2:2015 (E)
Table 1 — Summary of storage conditions for whole blood collection tubes with or without blood
genomic DNA stabilizers
Blood collection tube Blood Storage Analytical test type
Duration Temperature
With blood genomic a According to blood
— Blood collection tube manufacturer’s instructions
DNA stabilizer collection tube
a, b
— Analytical test providers’ instructions
manufacturer’s
instructions.
Without blood genomic d d Tests requiring
— ≤ 1 day — Room temperature
DNA stabilizer HMW DNA
— Analytical test — Analytical test
c c
instructions instructions
d d Tests requiring
— ≤ 3 days — 2 °C to 8 °C
HMW DNA
— Analytical test — Analytical test
c c
instructions instructions
d d Tests requiring
— ≤ 1 month — −20 °C
HMW DNA
— Analytical test — Analytical test
c c
instructions instructions
d d Tests not requiring
— ≤ 3 days — Room temperature
HMW DNA
— Analytical test — Analytical test
c c
instructions instructions
d d Tests not requiring
— ≤ 7 days — 2 °C to 8 °C
HMW DNA
— Analytical test — Analytical test
c c
instructions instructions
d d Tests not requiring
— ≤ 3 month — −20 °C
HMW DNA
— Analytical test — Analytical test
c c
instructions instructions
d d For all analytical tests
— Longer storage — −70 °C or below
— Analytical test — Analytical test
c c
instructions instructions
a
Requirement according to 5.1.4.2.
b
Alternative more stringent requirement according to 5.1.4.2.
c
Requirement according to 5.1.4.3.1.
d
Recommendation according to 5.1.4.3.2.
6.3 Isolation of the genomic DNA
6.3.1 General
The laboratory shall validate the entire process comprising sample collection, storage, transportation
and isolation of genomic DNA to the final analytical test result according to its internal quality
management system (see EN ISO 15189).
11
---------------------- Page: 13 ----------------------
SIST-TS CEN/TS 16835-2:2015
CEN/TS 16835-2:2015 (E)
To avoid a cross contamination with amplified DNA, the isolation of the blood genomic DNA should not
be performed in the same area as the amplification and post-amplification, unless a closed system is
used.
The genomic DNA isolation procedure chosen shall fulfil the requirements and specifications of the
intended molecular analytical test (e.g. DNA quality and quantity, DNA concentration, DNA length). If
the analytical test requires high molecular weight DNA, dedicated genomic DNA isolation methods such
as DNA precipitation-based methods should be applied [12]
The reagents and consumables coming in contact with the DNA sample shall be DNase-free.
6.3.2 Using commercial kits
1. When using commercial kits for the isolation of genomic DNA from blood, the manufacturers'
instructions for use shall be followed, unless 6.3.2, 4. and / or 6.3.3.1 apply.
2. When processing blood from tubes dedicated for genomic DNA testing and dedicated DNA isolation
kits are specified by the manufacturer of the blood collection tube, then these should be used for
the isolation of genomic DNA. Alternative extraction procedures can be used, if verified for the
same requirements and validated for the same intended use (6.3.3.1, 6.3.3.2).
3. When processing blood from tubes not dedicated for genomic DNA testing, commercially available
generic genomic DNA isolation kits can be used.
4. If the specifications of the analytical test provider require the use of a dedicated commercially
available kit, then this shall be used in accordance to the analytical test providers' instructions.
NOTE Dedicated procedures for use can be included in the manufacturers’ instructions for processing frozen
blood samples.
6.3.3 Using the laboratories own protocols
6.3.3.1 If a commercial kit is not used in accordance to its intended use, but is validate
...
Questions, Comments and Discussion
Ask us and Technical Secretary will try to provide an answer. You can facilitate discussion about the standard in here.