Molecular in vitro diagnostic examinations - Specifications for pre-examination processes for saliva - Isolated human DNA

This document gives requirements on the handling, storage, processing and documentation of saliva specimens intended for human DNA examination during the pre-examination phase before a molecular examination is performed.
This document is applicable to molecular in vitro diagnostic examination including laboratory developed tests performed by medical laboratories. It is also intended to be used by laboratory customers, in vitro diagnostics developers and manufacturers, biobanks, institutions and commercial organisations performing biomedical research, and regulatory authorities.
Dedicated measures that need to be taken for saliva collected on absorbing material or by mouth washes are not described in this technical specification. Neither are measures for preserving and handling of native saliva cell-free DNA, pathogens, and other bacterial or whole microbiome DNA in saliva described.
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 Speichel - Isolierte menschliche DNA

Analyses de diagnostic moléculaire in vitro - Spécifications relatives aux processus préanalytiques pour la salive - ADN humain extrait

Molekularne diagnostične preiskave in vitro - Specifikacije za predpreiskovalne procese za slino - Izolirana človeška DNK

Ta dokument podaja zahteve glede obravnave, shranjevanja, obdelave in dokumentiranja vzorcev sline, namenjenih za analizo DNK med predpreiskovalno fazo, preden se izvede molekularna preiskava.
Ta dokument se uporablja za molekularne diagnostične preiskave in vitro, vključno z laboratorijsko razvitimi preskusi, 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, in regulativni organi.
V tej tehnični specifikaciji niso opisani namenski ukrepi za slino, ki je zbrana na absorbcijskem materialu ali z ustno vodo. Prav tako niso opisani ukrepi za ohranjanje in ravnanje s slini lastnim brezceličnim DNK-jem, patogeni in drugim bakterijskim ali mikrobiomskim DNK-jem v slini.
OPOMBA:   Za določene teme, ki so zajete v tem dokumentu, lahko veljajo tudi mednarodni, nacionalni ali regionalni predpisi ali zahteve.

General Information

Status
Withdrawn
Publication Date
02-Apr-2019
Current Stage
9960 - Withdrawal effective - Withdrawal
Completion Date
03-Nov-2021

RELATIONS

Buy Standard

Technical specification
-TS CEN/TS 17305:2019
English language
17 pages
sale 10% off
Preview
sale 10% off
Preview
e-Library read for
1 day
Technical specification
-TS CEN/TS 17305:2019
English language
17 pages
sale 10% off
Preview
sale 10% off
Preview
e-Library read for
1 day

Standards Content (sample)

SLOVENSKI STANDARD
SIST-TS CEN/TS 17305:2019
01-julij-2019
Molekularne diagnostične preiskave in vitro - Specifikacije za predpreiskovalne
procese za slino - Izolirana človeška DNK

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

for saliva - Isolated human DNA
Molekularanalytische in-vitro-diagnostische Verfahren - Spezifikationen für
präanalytische Prozesse für Saliva - Isolierte menschliche DNS

Analyses de diagnostic moléculaire in vitro - Spécifications relatives aux processus

préanalytiques pour la salive - ADN humain isolé
Ta slovenski standard je istoveten z: CEN/TS 17305:2019
ICS:
11.100.10 Diagnostični preskusni In vitro diagnostic test
sistemi in vitro systems
SIST-TS CEN/TS 17305:2019 en,fr,de

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

---------------------- Page: 1 ----------------------
SIST-TS CEN/TS 17305:2019
---------------------- Page: 2 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305
TECHNICAL SPECIFICATION
SPÉCIFICATION TECHNIQUE
April 2019
TECHNISCHE SPEZIFIKATION
ICS 11.100.10
English Version
Molecular in vitro diagnostic examinations - Specifications
for pre-examination processes for saliva - Isolated human
DNA

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

Spécifications relatives aux processus préanalytiques - Spezifikationen für präanalytische Prozesse für

pour la salive - ADN humain extrait Speichel - Isolierte menschliche DNA

This Technical Specification (CEN/TS) was approved by CEN on 21 January 2019 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, 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. CEN/TS 17305:2019 E

worldwide for CEN national Members.
---------------------- Page: 3 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305:2019 (E)
Contents Page

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

Introduction .................................................................................................................................................................... 4

1 Scope .................................................................................................................................................................... 5

2 Normative references .................................................................................................................................... 5

3 Terms and definitions ................................................................................................................................... 5

4 General Considerations................................................................................................................................. 9

5 Outside the laboratory ............................................................................................................................... 10

5.1 Specimen collection ..................................................................................................................................... 10

5.1.1 Information about the specimen donor/patient .............................................................................. 10

5.1.2 Selection of the saliva collection device by the laboratory ........................................................... 10

5.1.3 Saliva specimen collection from the donor/patient and stabilization procedures ............. 10

5.1.4 Information on the specimen and storage requirements at saliva collection

facility/site ..................................................................................................................................................... 11

5.2 Transport requirements............................................................................................................................ 12

5.2.1 General ............................................................................................................................................................. 12

5.2.2 Using saliva collection devices with DNA stabilizers ...................................................................... 12

5.2.3 Using saliva collection devices without DNA stabilizers ................................................................ 12

6 Inside the laboratory .................................................................................................................................. 12

6.1 Specimen reception ..................................................................................................................................... 12

6.2 Storage requirements ................................................................................................................................. 13

6.3 Isolation of the saliva DNA ........................................................................................................................ 13

6.3.1 General ............................................................................................................................................................. 13

6.3.2 Using commercial kit .................................................................................................................................. 13

6.3.3 Using the laboratories own protocol .................................................................................................... 14

6.4 Quantity and quality assessment of isolated DNA ............................................................................ 14

6.5 Storage of isolated saliva DNA ................................................................................................................. 14

6.5.1 General ............................................................................................................................................................. 14

6.5.2 Saliva DNA isolated with commercially available kits .................................................................... 15

6.5.3 Saliva DNA isolated with the laboratory's own protocols ............................................................. 15

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

---------------------- Page: 4 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305:2019 (E)
European foreword

This document (CEN/TS 17305:2019) 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.

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, Former Yugoslav Republic of Macedonia,

France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta,

Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,

Turkey and the United Kingdom.
---------------------- Page: 5 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305:2019 (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

specimen 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.

Genetic examination of DNA is commonly used in clinical practice. This includes e.g. predisposition

testing, pharmacogenomics, analysis of genetic disorders with the perspective use in precision

medicine. This is a fast growing field in molecular diagnostics.

Saliva is increasingly used as a non-invasive alternative specimen to blood for the examination of

human DNA. Saliva naturally contains microorganisms and also extraneous substances (e.g., food

debris), which make the composition of saliva more complex and unique among patients/donors.

Dedicated measures are therefore needed for informing and preparing patients/donors for the

collection and to check compliance with the instructions, in order to reduce the specimen variability. In

contrast to invasive specimen collection, saliva collection does not require trained and educated

professionals or dedicated facilities. By good instruction and verified collection device safety claims,

saliva specimens can be self-collected at home; however, home collection also contributes to high

variability in specimen quality. Similarly, medical laboratories/ in vitro manufacturers need to be aware

of specimen variability when performing design verification and validation.

DNA in saliva can fragment or degrade after collection. In addition, bacteria present in the saliva

specimen can continue to grow, thus diluting the human DNA. DNases secreted by these bacteria can

also accelerate the DNA degradation. This can impact the sensitivity and reliability of DNA examination.

Standardization of the entire process from specimen collection to the DNA examination is needed to

minimize DNA degradation and fragmentation after saliva collection. This document describes special

measures which need to be taken to obtain good quality saliva specimen/samples and isolated DNA

therefrom for human DNA examination.
In this document, the following verbal forms are used:
— “shall” indicates a requirement;
— “should” indicates a recommendation;
— “may” indicates a permission;
— “can” indicates a possibility or a capability.
---------------------- Page: 6 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305:2019 (E)
1 Scope

This document gives requirements on the handling, storage, processing and documentation of saliva

specimens intended for human DNA examination during the pre-examination phase before a molecular

examination is performed.

This document is applicable to molecular in vitro diagnostic examination including laboratory

developed tests performed by medical laboratories. It is also intended to be used by laboratory

customers, in vitro diagnostics developers and manufacturers, biobanks, institutions and commercial

organisations performing biomedical research, and regulatory authorities.

Dedicated measures that need to be taken for saliva collected on absorbing material or by mouth

washes are not described in this technical specification. Neither are measures for preserving and

handling of native saliva cell-free DNA, pathogens, and other bacterial or whole microbiome DNA in

saliva described.

NOTE International, national or regional regulations or requirements can also apply to specific topics

covered in this document.
2 Normative references

The following documents are referred to in the text in such a way that some or all of their content

constitutes requirements of this document. For dated references, only the edition cited applies. For

undated references, the latest edition of the referenced document (including any amendments) applies.

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 and the following

apply.

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

• IEC Electropedia: available at http://www.electropedia.org/
• ISO Online browsing platform: available at http://www.iso.org/obp
3.1
ambient temperature
unregulated temperature of the surrounding air
3.2
analyte
component represented in the name of a measurable quantity
[SOURCE: ISO 17511:2003, 3.2 modified — The examples were not taken over.]
---------------------- Page: 7 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305:2019 (E)
3.3
examination performance
analytical test performance
analytical performance

accuracy, precision, and sensitivity of a test to measure the analyte of interest

Note 1 to entry: Other test performance characteristics such as robustness, repeatability can apply as well.

[SOURCE: ISO 20184-1:2018, 3.4]
3.4
DNA stabilizers

compounds, solutions or mixtures that are designed to minimize degradation and fragmentation of DNA

3.5
closed system

non-modifiable system provided by the vendor including all necessary components for the analysis (i.e.,

hardware, software, procedures and reagents)
3.6
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]
3.7
DNA proficiency testing program
proficiency testing for DNA based examinations

Note 1 to entry: Commonly, a program periodically sends multiple specimens to members of a group of

laboratories for analysis and/or identification; the program then compares each laboratory’s results with those of

other laboratories in the group and/or with an assigned value, and reports the results to the participating

laboratory and others.

Note 2 to entry: Other forms of PT/EQA include: data transformation exercises, single-item testing (where one

item is sent to a number of laboratories sequentially and returned to the program at intervals), and one-off

exercises (where laboratories are provided with a test item on a single occasion).

3.8
examination
analytical test

set of operations having the object of determining the value or characteristics of a property

[SOURCE: EN ISO 15189:2012, 3.7, modified — The term and definition is used here without the

original notes.]

Note 1 to entry: Processes that start with the isolated measurand and include all kinds of parameter testing or

chemical manipulation for quantitative or qualitative examination.
3.9
examination provider
analytical test provider
group or company that provides the specific analytical test
---------------------- Page: 8 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305:2019 (E)
3.10
interfering substances

endogenous or exogenous substances (e.g. stabilization solution) that can be present in specimens and

that can alter an examination result
3.11
microorganism
entity of microscopic size, encompassing bacteria, fungi, protozoa and viruses
[SOURCE: ISO 11139:2018, 3.176]
3.12
pre-examination processes
pre-analytical phase
pre-analytical 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 analytical laboratory, 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 detail was

included.]

Note 1 to entry: The pre-examination phase includes preparative processes that influence the outcome of the

intended examination.
3.13
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.]
3.14
proficiency testing

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

comparisons

[SOURCE: ISO/IEC 17043:2010, 3.7, modified — Term and definition are used here without the original

notes.]
3.15
room temperature
for the purpose of this document, temperature in the range of 18 °C to 25 °C
Note 1 to entry: Local or national regulations can have different definitions.
3.16
saliva
whole saliva

bio-fluid of the mouth composed mainly of secretion originating from the three major salivary glands

(parotids, submandibular and sublingual glands) and from salivary glands present in the oral cavity

---------------------- Page: 9 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305:2019 (E)
3.17
saliva collection device
tube or other container in which the saliva specimen is collected
3.18
sample
one or more parts taken from a primary sample
[SOURCE: EN ISO 15189:2012, 3.24, modified — The examples were not taken over.]
3.19
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

Note 1 to entry: The measurand constituent for the purpose of this document is isolated DNA.

[SOURCE ISO Guide 30:2015, 2.1.15, modified — Note 1 was not taken over. The following words

were replaced: “characteristic” by “ability”; “reference material” by “sample material”; “specified” by

“stated”.]
3.20
storage

prolonged interruption of the pre-analytical workflow of a sample or analyte respectively, or of their

derivatives, under appropriate conditions in order to preserve their properties

Note 1 to entry: Long-term storage typically occurs in laboratory archives or in biobanks.

[SOURCE: ISO 20184-1:2018, 3.22, modified — Example in the definition was deleted.]

3.21
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 word “validated” is used to designate the corresponding status.

[SOURCE: EN ISO 9000:2015, 3.8.13, modified — Note 1 and 3 were not taken over.]
---------------------- Page: 10 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305:2019 (E)
3.22
verification

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

Note 1 to entry: The word “verified” is used to designate the corresponding status.

[SOURCE: EN ISO 9000:2015, 3.8.12, modified — Note 1 and Note 2 were not taken over.]

Note 2 to entry: Confirmation can comprise activities such as
— performing alternative calculations;

— comparing a new design specification with a similar proven design specification;

— undertaking tests and demonstrations; and
— reviewing documents prior to issue.
3.23
workflow
series of activities necessary to complete a task
4 General Considerations

For general statements on medical laboratory quality management systems and in particular on

primary sample collection, reception and handling (including avoidance of cross contaminations) see

EN ISO 15189:2012, 4.2 and 5.4.4, 5.4.6, or EN ISO/IEC 17020:2012, Clause 8 and 7.2. The requirements

on laboratory equipment, reagents, and consumables according to EN ISO 15189:2012, 5.3 shall be

followed; EN ISO 15189:2012, 5.5.1.2 and 5.5.1.3, and EN 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 their impact on the stability of

biomolecules intended to be examined shall be verified and validated for its intended use. The stability

of the human DNA should be investigated throughout the complete pre-examination process

development. The verification of performance claims as well as the validation of the examination shall

take into account the variability of the saliva specimen's quality. A risk assessment of non-controllable

workflow steps including their potential impact on the examination performance shall be performed

and mitigation measures shall be established to enable the required examination performance. Before

or during the design of an examination, it therefore should be investigated and ensured that the human

DNA minimum amount and size required for the examination is/are not compromized in a manner

impacting the examination performance.

Safety regulations on facilities, transport and handling shall be considered (EN ISO 15189:2012, 5.2.3

and 5.4.5, and ISO 15190). Safety precautions according to ISO 15190 shall be followed.

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

between different specimens/samples,
...

SLOVENSKI STANDARD
SIST-TS CEN/TS 17305:2019
01-julij-2019
Molekularne diagnostične preiskave in vitro - Specifikacije za predpreiskovalne
procese za slino - Izolirani človeški DNK

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

for saliva - Isolated human DNA
Molekularanalytische in-vitro-diagnostische Verfahren - Spezifikationen für
präanalytische Prozesse für Saliva - Isolierte menschliche DNS

Analyses de diagnostic moléculaire in vitro - Spécifications relatives aux processus

préanalytiques pour la salive - ADN humain isolé
Ta slovenski standard je istoveten z: CEN/TS 17305:2019
ICS:
11.100.10 Diagnostični preskusni In vitro diagnostic test
sistemi in vitro systems
SIST-TS CEN/TS 17305:2019 en,fr,de

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

---------------------- Page: 1 ----------------------
SIST-TS CEN/TS 17305:2019
---------------------- Page: 2 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305
TECHNICAL SPECIFICATION
SPÉCIFICATION TECHNIQUE
April 2019
TECHNISCHE SPEZIFIKATION
ICS 11.100.10
English Version
Molecular in vitro diagnostic examinations - Specifications
for pre-examination processes for saliva - Isolated human
DNA

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

Spécifications relatives aux processus préanalytiques - Spezifikationen für präanalytische Prozesse für

pour la salive - ADN humain extrait Speichel - Isolierte menschliche DNA

This Technical Specification (CEN/TS) was approved by CEN on 21 January 2019 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, 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. CEN/TS 17305:2019 E

worldwide for CEN national Members.
---------------------- Page: 3 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305:2019 (E)
Contents Page

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

Introduction .................................................................................................................................................................... 4

1 Scope .................................................................................................................................................................... 5

2 Normative references .................................................................................................................................... 5

3 Terms and definitions ................................................................................................................................... 5

4 General Considerations................................................................................................................................. 9

5 Outside the laboratory ............................................................................................................................... 10

5.1 Specimen collection ..................................................................................................................................... 10

5.1.1 Information about the specimen donor/patient .............................................................................. 10

5.1.2 Selection of the saliva collection device by the laboratory ........................................................... 10

5.1.3 Saliva specimen collection from the donor/patient and stabilization procedures ............. 10

5.1.4 Information on the specimen and storage requirements at saliva collection

facility/site ..................................................................................................................................................... 11

5.2 Transport requirements............................................................................................................................ 12

5.2.1 General ............................................................................................................................................................. 12

5.2.2 Using saliva collection devices with DNA stabilizers ...................................................................... 12

5.2.3 Using saliva collection devices without DNA stabilizers ................................................................ 12

6 Inside the laboratory .................................................................................................................................. 12

6.1 Specimen reception ..................................................................................................................................... 12

6.2 Storage requirements ................................................................................................................................. 13

6.3 Isolation of the saliva DNA ........................................................................................................................ 13

6.3.1 General ............................................................................................................................................................. 13

6.3.2 Using commercial kit .................................................................................................................................. 13

6.3.3 Using the laboratories own protocol .................................................................................................... 14

6.4 Quantity and quality assessment of isolated DNA ............................................................................ 14

6.5 Storage of isolated saliva DNA ................................................................................................................. 14

6.5.1 General ............................................................................................................................................................. 14

6.5.2 Saliva DNA isolated with commercially available kits .................................................................... 15

6.5.3 Saliva DNA isolated with the laboratory's own protocols ............................................................. 15

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

---------------------- Page: 4 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305:2019 (E)
European foreword

This document (CEN/TS 17305:2019) 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.

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, Former Yugoslav Republic of Macedonia,

France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta,

Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,

Turkey and the United Kingdom.
---------------------- Page: 5 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305:2019 (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

specimen 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.

Genetic examination of DNA is commonly used in clinical practice. This includes e.g. predisposition

testing, pharmacogenomics, analysis of genetic disorders with the perspective use in precision

medicine. This is a fast growing field in molecular diagnostics.

Saliva is increasingly used as a non-invasive alternative specimen to blood for the examination of

human DNA. Saliva naturally contains microorganisms and also extraneous substances (e.g., food

debris), which make the composition of saliva more complex and unique among patients/donors.

Dedicated measures are therefore needed for informing and preparing patients/donors for the

collection and to check compliance with the instructions, in order to reduce the specimen variability. In

contrast to invasive specimen collection, saliva collection does not require trained and educated

professionals or dedicated facilities. By good instruction and verified collection device safety claims,

saliva specimens can be self-collected at home; however, home collection also contributes to high

variability in specimen quality. Similarly, medical laboratories/ in vitro manufacturers need to be aware

of specimen variability when performing design verification and validation.

DNA in saliva can fragment or degrade after collection. In addition, bacteria present in the saliva

specimen can continue to grow, thus diluting the human DNA. DNases secreted by these bacteria can

also accelerate the DNA degradation. This can impact the sensitivity and reliability of DNA examination.

Standardization of the entire process from specimen collection to the DNA examination is needed to

minimize DNA degradation and fragmentation after saliva collection. This document describes special

measures which need to be taken to obtain good quality saliva specimen/samples and isolated DNA

therefrom for human DNA examination.
In this document, the following verbal forms are used:
— “shall” indicates a requirement;
— “should” indicates a recommendation;
— “may” indicates a permission;
— “can” indicates a possibility or a capability.
---------------------- Page: 6 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305:2019 (E)
1 Scope

This document gives requirements on the handling, storage, processing and documentation of saliva

specimens intended for human DNA examination during the pre-examination phase before a molecular

examination is performed.

This document is applicable to molecular in vitro diagnostic examination including laboratory

developed tests performed by medical laboratories. It is also intended to be used by laboratory

customers, in vitro diagnostics developers and manufacturers, biobanks, institutions and commercial

organisations performing biomedical research, and regulatory authorities.

Dedicated measures that need to be taken for saliva collected on absorbing material or by mouth

washes are not described in this technical specification. Neither are measures for preserving and

handling of native saliva cell-free DNA, pathogens, and other bacterial or whole microbiome DNA in

saliva described.

NOTE International, national or regional regulations or requirements can also apply to specific topics

covered in this document.
2 Normative references

The following documents are referred to in the text in such a way that some or all of their content

constitutes requirements of this document. For dated references, only the edition cited applies. For

undated references, the latest edition of the referenced document (including any amendments) applies.

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 and the following

apply.

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

• IEC Electropedia: available at http://www.electropedia.org/
• ISO Online browsing platform: available at http://www.iso.org/obp
3.1
ambient temperature
unregulated temperature of the surrounding air
3.2
analyte
component represented in the name of a measurable quantity
[SOURCE: ISO 17511:2003, 3.2 modified — The examples were not taken over.]
---------------------- Page: 7 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305:2019 (E)
3.3
examination performance
analytical test performance
analytical performance

accuracy, precision, and sensitivity of a test to measure the analyte of interest

Note 1 to entry: Other test performance characteristics such as robustness, repeatability can apply as well.

[SOURCE: ISO 20184-1:2018, 3.4]
3.4
DNA stabilizers

compounds, solutions or mixtures that are designed to minimize degradation and fragmentation of DNA

3.5
closed system

non-modifiable system provided by the vendor including all necessary components for the analysis (i.e.,

hardware, software, procedures and reagents)
3.6
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]
3.7
DNA proficiency testing program
proficiency testing for DNA based examinations

Note 1 to entry: Commonly, a program periodically sends multiple specimens to members of a group of

laboratories for analysis and/or identification; the program then compares each laboratory’s results with those of

other laboratories in the group and/or with an assigned value, and reports the results to the participating

laboratory and others.

Note 2 to entry: Other forms of PT/EQA include: data transformation exercises, single-item testing (where one

item is sent to a number of laboratories sequentially and returned to the program at intervals), and one-off

exercises (where laboratories are provided with a test item on a single occasion).

3.8
examination
analytical test

set of operations having the object of determining the value or characteristics of a property

[SOURCE: EN ISO 15189:2012, 3.7, modified — The term and definition is used here without the

original notes.]

Note 1 to entry: Processes that start with the isolated measurand and include all kinds of parameter testing or

chemical manipulation for quantitative or qualitative examination.
3.9
examination provider
analytical test provider
group or company that provides the specific analytical test
---------------------- Page: 8 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305:2019 (E)
3.10
interfering substances

endogenous or exogenous substances (e.g. stabilization solution) that can be present in specimens and

that can alter an examination result
3.11
microorganism
entity of microscopic size, encompassing bacteria, fungi, protozoa and viruses
[SOURCE: ISO 11139:2018, 3.176]
3.12
pre-examination processes
pre-analytical phase
pre-analytical 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 analytical laboratory, 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 detail was

included.]

Note 1 to entry: The pre-examination phase includes preparative processes that influence the outcome of the

intended examination.
3.13
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.]
3.14
proficiency testing

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

comparisons

[SOURCE: ISO/IEC 17043:2010, 3.7, modified — Term and definition are used here without the original

notes.]
3.15
room temperature
for the purpose of this document, temperature in the range of 18 °C to 25 °C
Note 1 to entry: Local or national regulations can have different definitions.
3.16
saliva
whole saliva

bio-fluid of the mouth composed mainly of secretion originating from the three major salivary glands

(parotids, submandibular and sublingual glands) and from salivary glands present in the oral cavity

---------------------- Page: 9 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305:2019 (E)
3.17
saliva collection device
tube or other container in which the saliva specimen is collected
3.18
sample
one or more parts taken from a primary sample
[SOURCE: EN ISO 15189:2012, 3.24, modified — The examples were not taken over.]
3.19
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

Note 1 to entry: The measurand constituent for the purpose of this document is isolated DNA.

[SOURCE ISO Guide 30:2015, 2.1.15, modified — Note 1 was not taken over. The following words

were replaced: “characteristic” by “ability”; “reference material” by “sample material”; “specified” by

“stated”.]
3.20
storage

prolonged interruption of the pre-analytical workflow of a sample or analyte respectively, or of their

derivatives, under appropriate conditions in order to preserve their properties

Note 1 to entry: Long-term storage typically occurs in laboratory archives or in biobanks.

[SOURCE: ISO 20184-1:2018, 3.22, modified — Example in the definition was deleted.]

3.21
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 word “validated” is used to designate the corresponding status.

[SOURCE: EN ISO 9000:2015, 3.8.13, modified — Note 1 and 3 were not taken over.]
---------------------- Page: 10 ----------------------
SIST-TS CEN/TS 17305:2019
CEN/TS 17305:2019 (E)
3.22
verification

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

Note 1 to entry: The word “verified” is used to designate the corresponding status.

[SOURCE: EN ISO 9000:2015, 3.8.12, modified — Note 1 and Note 2 were not taken over.]

Note 2 to entry: Confirmation can comprise activities such as
— performing alternative calculations;

— comparing a new design specification with a similar proven design specification;

— undertaking tests and demonstrations; and
— reviewing documents prior to issue.
3.23
workflow
series of activities necessary to complete a task
4 General Considerations

For general statements on medical laboratory quality management systems and in particular on

primary sample collection, reception and handling (including avoidance of cross contaminations) see

EN ISO 15189:2012, 4.2 and 5.4.4, 5.4.6, or EN ISO/IEC 17020:2012, Clause 8 and 7.2. The requirements

on laboratory equipment, reagents, and consumables according to EN ISO 15189:2012, 5.3 shall be

followed; EN ISO 15189:2012, 5.5.1.2 and 5.5.1.3, and EN 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 their impact on the stability of

biomolecules intended to be examined shall be verified and validated for its intended use. The stability

of the human DNA should be investigated throughout the complete pre-examination process

development. The verification of performance claims as well as the validation of the examination shall

take into account the variability of the saliva specimen's quality. A risk assessment of non-controllable

workflow steps including their potential impact on the examination performance shall be performed

and mitigation measures shall be established to enable the required examination performance. Before

or during the design of an examination, it therefore should be investigated and ensured that the human

DNA minimum amount and size required for the examination is/are not compromized in a manner

impacting the examination performance.

Safety regulations on facilities, transport and handling shall be considered (EN ISO 15189:2012, 5.2.3

and 5.4.5, and ISO 15190). Safety precautions according to ISO 15190 shall be followed.

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

between different specimens/samples,
...

Questions, Comments and Discussion

Ask us and Technical Secretary will try to provide an answer. You can facilitate discussion about the standard in here.