SIST EN ISO 17099:2024
(Main)Radiological protection - Performance criteria for laboratories using the cytokinesis block micronucleus (CBMN) assay in peripheral blood lymphocytes for biological dosimetry (ISO 17099:2024)
Radiological protection - Performance criteria for laboratories using the cytokinesis block micronucleus (CBMN) assay in peripheral blood lymphocytes for biological dosimetry (ISO 17099:2024)
This document gives guidance on
a) confidentiality of personal information for the customer and the laboratory,
b) laboratory safety requirements,
c) calibration sources and calibration dose ranges useful for establishing the reference dose-response curves that contribute to the dose estimation from CBMN assay yields and the detection limit,
d) performance of blood collection, culturing, harvesting, and sample preparation for CBMN assay scoring,
e) scoring criteria,
f) conversion of micronucleus frequency in BNCs into an estimate of absorbed dose,
g) reporting of results,
h) quality assurance and quality control, and
i) informative annexes containing sample instructions for customers, sample questionnaire, a microscope scoring data sheet, and a sample report.
This document excludes methods for automated scoring of CBMN.
Strahlenschutz - Leistungskriterien für Laboratorien, die den Zytokineseblock-Mikronukleustest (CBMN) in peripheren Blutlymphozyten für die biologische Dosimetrie verwenden (ISO 17099:2024)
Dieses Dokument behandelt Folgendes:
a) die Vertraulichkeit von persönlichen Angaben, in Bezug auf Auftraggeber und Laboratorium;
b) die Labor-Sicherheitsanforderungen;
c) die für die Bestimmung der Referenz-Dosis-Effekt-Kurven verwendeten Kalibrierungsquellen und dosisbereiche, mit denen aus den Ergebnissen des CBMN-Tests und der Nachweisgrenze die Dosis geschätzt wird;
d) die Durchführung der Blutentnahme, Kultivierung, Aufarbeitung und Probenpräparation für den CBMN-Test;
e) die Auswertekriterien;
f) die Abschätzung der absorbierten Dosis anhand der Häufigkeit von Mikrokernen in BNCs;
g) die Berichterstattung der Ergebnisse;
h) Qualitätssicherung und Qualitätskontrolle, und
i) informative Anhänge mit Musteranweisungen für Auftraggeber, Musterfragebogen, Musterdatenblatt für die mikroskopische Auswertung und Musterbericht.
Dieses Dokument schließt Verfahren zur automatischen Bewertung von CBMN aus.
Radioprotection - Critères de performance pour les laboratoires pratiquant la dosimétrie biologique par l’analyse des micronoyaux par blocage de la cytodiérèse (CBMN) dans les lymphocytes du sang périphérique (ISO 17099:2024)
Le présent document fournit des recommandations relatives:
a) à la confidentialité des informations personnelles pour le demandeur et le laboratoire;
b) aux exigences de sécurité relatives aux laboratoires;
c) aux sources d’étalonnage et aux gammes de doses d’étalonnage utiles pour établir les courbes dose‑effet de référence qui contribuent à l’estimation de dose à partir des résultats de l’analyse des CBMN et à la limite de détection;
d) au prélèvement, à la mise en culture, au recueil des cellules cultivées et à la préparation des échantillons de sang pour le dénombrement par analyse des CBMN;
e) aux résultats de dénombrement;
f) à la conversion de la fréquence des micronoyaux dans les BNC en estimation de dose absorbée;
g) à la présentation des résultats;
h) à l’assurance qualité et au contrôle qualité;
i) aux annexes informatives contenant des exemples d’instructions d’utilisation, de questionnaire, de tableau de dénombrement au microscope et de rapport.
Le présent document exclut les méthodes de dénombrement automatisé des CBMN.
Radiološka zaščita - Merila za delovanje laboratorijev, ki za biološko dozimetrijo uporabljajo analizo tvorjenja mikrojeder s citokinetskim blokom (CBMN) v perifernih krvnih limfocitih (ISO 17099:2024)
Ta dokument podaja smernice za
a) zaupnost osebnih podatkov stranke in laboratorija,
b) zahteve za varnost laboratorija,
c) vire umerjanja in razpone odmerkov umerjanja, ki so koristni za določanje referenčnih krivulj odziva odmerka ter omogočajo ocenjevanje odmerka na podlagi rezultatov analize tvorjenja mikrojeder s citokinetskim blokom (CBMN) in meje zaznavnosti,
d) učinkovitost zbiranja krvi ter gojenja, zajemanja in priprave vzorcev za ocenjevanje rezultatov analize tvorjenja mikrojeder s citokinetskim blokom,
e) kriterije ocenjevanja,
f) pretvorbo pogostosti mikrojeder v dvojedrnih celicah (BNC) v približek absorbiranega odmerka,
g) poročanje rezultatov,
h) zagotavljanje in nadzor kakovosti ter
i) informativne dodatke, ki vsebujejo vzorčna navodila za stranke, vzorčni vprašalnik, tehnični list za ocenjevanje pri opazovanju z mikroskopom in vzorčno poročilo.
Ta dokument ne zajema metod za avtomatizirano ocenjevanje tvorjenja mikrojeder s citokinetskim blokom.
General Information
Relations
Overview
EN ISO 17099:2024 - Radiological protection - Performance criteria for laboratories using the cytokinesis-block micronucleus (CBMN) assay in peripheral blood lymphocytes for biological dosimetry - provides performance guidance for laboratories that use the CBMN assay to estimate absorbed radiation dose from micronucleus yields in peripheral blood lymphocytes. Endorsed by CEN in 2024, this standard replaces EN ISO 17099:2017 and defines technical, quality and reporting expectations for biological dosimetry using the CBMN method. Note: methods for automated scoring of CBMN are excluded.
Key topics and requirements
- Confidentiality and data protection: handling and storage of personal information, anonymity of samples, transmission and reporting of confidential results.
- Laboratory safety: microbiological, chemical and optical safety measures and an overarching safety plan.
- Calibration and sensitivity: recommended calibration sources, calibration dose ranges to establish reference dose–response curves, background micronucleus (MN) frequency, and methods to characterise the minimum detectable dose.
- Sample processing and scoring: procedures for blood collection, culturing, harvesting, staining, microscopy and clear scoring criteria for micronuclei in binucleated cells (BNCs).
- Dose estimation: converting MN frequency in BNCs to an absorbed dose estimate, including calculations, uncertainty, comparison with controls and handling acute vs non‑acute exposures.
- Quality assurance / quality control (QA/QC): requirements for setting up and sustaining QA programs and routine QC measures.
- Reporting: content and format of result reports, interpretation guidance and practical annex material.
- Annexes: informative examples such as customer instructions, sample questionnaires, a microscope scoring data sheet and a sample report.
Practical applications
- Rapid biological dose assessment after accidental or suspected radiation exposure (individual or population triage).
- Support for emergency response teams, occupational health, radiation protection services and public health agencies assessing exposed individuals.
- Standardization of inter-laboratory comparisons and networks for large-scale event response.
- Ensuring consistent QA/QC and traceable dose estimation for medico-legal, clinical and regulatory reporting.
Who should use this standard
- Diagnostic and service laboratories performing CBMN biodosimetry
- Radiation protection specialists and emergency response labs
- Public health authorities, occupational health providers and research groups working on biological dosimetry
Related standards (applicable)
- ISO 17099:2024 is complementary to laboratory competence and quality standards (e.g., ISO/IEC 17025 for testing laboratories) for establishing validated, auditable biodosimetry services.
Keywords: EN ISO 17099:2024, CBMN assay, cytokinesis-block micronucleus, biological dosimetry, radiological protection, peripheral blood lymphocytes, calibration curve, minimum detectable dose, micronucleus frequency, binucleated cells, laboratory QA/QC.
Standards Content (Sample)
SLOVENSKI STANDARD
01-september-2024
Radiološka zaščita - Merila za delovanje laboratorijev, ki za biološko dozimetrijo
uporabljajo analizo tvorjenja mikrojeder s citokinetskim blokom (CBMN) v
perifernih krvnih limfocitih (ISO 17099:2024)
Radiological protection - Performance criteria for laboratories using the cytokinesis block
micronucleus (CBMN) assay in peripheral blood lymphocytes for biological dosimetry
(ISO 17099:2024)
Strahlenschutz - Leistungskriterien für Laboratorien, die den Zytokineseblock-
Mikronukleustest (CBMN) in peripheren Blutlymphozyten für die biologische Dosimetrie
verwenden (ISO 17099:2024)
Radioprotection - Critères de performance pour les laboratoires pratiquant la dosimétrie
biologique par l’analyse des micronoyaux par blocage de la cytodiérèse (CBMN) dans
les lymphocytes du sang périphérique (ISO 17099:2024)
Ta slovenski standard je istoveten z: EN ISO 17099:2024
ICS:
13.280 Varstvo pred sevanjem Radiation protection
71.040.10 Kemijski laboratoriji. Chemical laboratories.
Laboratorijska oprema Laboratory equipment
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EN ISO 17099
EUROPEAN STANDARD
NORME EUROPÉENNE
June 2024
EUROPÄISCHE NORM
ICS 13.280 Supersedes EN ISO 17099:2017
English Version
Radiological protection - Performance criteria for
laboratories using the cytokinesis-block micronucleus
(CBMN) assay in peripheral blood lymphocytes for
biological dosimetry (ISO 17099:2024)
Radioprotection - Critères de performance pour les Strahlenschutz - Leistungskriterien für Laboratorien,
laboratoires pratiquant la dosimétrie biologique par die den Zytokineseblock-Mikronukleustest (CBMN) in
l'analyse des micronoyaux par blocage de la peripheren Blutlymphozyten für die biologische
cytodiérèse (CBMN) dans les lymphocytes du sang Dosimetrie verwenden (ISO 17099:2024)
périphérique (ISO 17099:2024)
This European Standard was approved by CEN on 24 June 2024.
CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this
European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references
concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN
member.
This European Standard exists in three official versions (English, French, German). A version in any other language made by
translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management
Centre has the same status as the official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye 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
© 2024 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 17099:2024 E
worldwide for CEN national Members.
Contents Page
European foreword . 3
European foreword
This document (EN ISO 17099:2024) has been prepared by Technical Committee ISO/TC 85 "Nuclear
energy, nuclear technologies, and radiological protection" in collaboration with Technical Committee
CEN/TC 430 “Nuclear energy, nuclear technologies, and radiological protection” the secretariat of
which is held by AFNOR.
This European Standard shall be given the status of a national standard, either by publication of an
identical text or by endorsement, at the latest by December 2024, and conflicting national standards
shall be withdrawn at the latest by December 2024.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
This document supersedes EN ISO 17099:2017.
Any feedback and questions on this document should be directed to the users’ national standards
body/national committee. A complete listing of these bodies can be found on the CEN website.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the
following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria,
Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland,
Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Republic of
North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and the
United Kingdom.
Endorsement notice
The text of ISO 17099:2024 has been approved by CEN as EN ISO 17099:2024 without any modification.
International
Standard
ISO 17099
Second edition
Radiological protection —
2024-06
Performance criteria for
laboratories using the cytokinesis-
block micronucleus (CBMN) assay
in peripheral blood lymphocytes for
biological dosimetry
Radioprotection — Critères de performance pour les
laboratoires pratiquant la dosimétrie biologique par l’analyse
des micronoyaux par blocage de la cytodiérèse (CBMN) dans les
lymphocytes du sang périphérique
Reference number
ISO 17099:2024(en) © ISO 2024
ISO 17099:2024(en)
© ISO 2024
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on
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ISO copyright office
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Phone: +41 22 749 01 11
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii
ISO 17099:2024(en)
Contents Page
Foreword .v
Introduction .vii
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 CMBN assay methodology used in this document . 3
4.1 General .3
4.2 Requests for analysis and blood sampling .3
5 Responsibility of the requestor . 3
6 Responsibility of the service laboratory . 4
6.1 Setup and sustainment of the quality assurance program .4
6.2 Responsibility during service .4
7 Confidentiality of personal information . 5
7.1 Overview .5
7.2 Applications of the principle of confidentiality .5
7.2.1 Delegation of responsibilities within the laboratory.5
7.2.2 Requests for analysis . .6
7.2.3 Transmission of confidential information .6
7.2.4 Anonymity of samples .6
7.2.5 Reporting of results .6
7.2.6 Storage .6
7.2.7 Data security plan .6
8 Laboratory safety requirements . 6
8.1 Overview .6
8.2 Microbiological safety requirements .7
8.3 Chemical safety requirements .7
8.4 Optical safety requirements .8
8.5 Safety plan .8
9 Sample processing. 8
9.1 Culturing .8
9.2 Staining .10
9.3 Microscopy .10
9.4 Scoring of slides .10
9.4.1 General .10
9.4.2 Criteria for scoring .10
9.4.3 Scoring data sheets .11
9.5 Automated analysis .11
10 Calibration source(s), calibration curve, and minimum detectable dose.11
10.1 Calibration source(s) .11
10.2 Calibration curve .11
10.3 Background MN frequency . 12
10.4 Comparison with the background level: Characterisation of the minimum detectable
dose . 13
11 Accidental exposure involving few individuals .15
11.1 Procedure for scoring MN in BNCs . 15
11.1.1 Coding of samples and slides . 15
11.1.2 Scoring techniques .16
11.1.3 Laboratory scoring expertise .16
11.2 Criteria for converting a MN yield into an estimate of absorbed dose .16
11.2.1 Overview .16
iii
ISO 17099:2024(en)
11.2.2 Comparison with controls .16
11.2.3 Confidence limits on the number of MN .16
11.2.4 Calculation of absorbed dose for whole-body exposures .16
11.2.5 Calculation of uncertainty on absorbed dose .17
11.2.6 Acute and non-acute exposure cases .17
11.2.7 Testing the distribution of MN per BNC .18
11.2.8 Other exposure scenarios .18
11.3 Reporting of results .18
11.3.1 General .18
11.3.2 Content of the report (see Annex D for a standard form) .18
11.3.3 Interpretation of the results .19
12 Population triage . 19
12.1 General .19
12.2 Use of a CBMN assay network for large scale exposures .19
12.3 Procedure for scoring MN in BNCs . 20
12.4 Criteria for converting a MN yield into an estimate of absorbed dose . 20
12.5 Reporting of results . 20
13 Quality assurance and quality control .20
13.1 Overview . 20
13.2 Specific requirement . 20
13.2.1 General . 20
13.2.2 Performance checks by laboratory inter-comparisons . 20
13.2.3 Periodical performance check of scorer qualification .21
13.2.4 Performance checks of sample transport integrity .21
13.2.5 Performance checks of sample integrity by service laboratory .21
13.2.6 Performance checks for instrumentation .21
13.2.7 Performance checks of sample protocol . 22
13.2.8 Performance checks of sample scoring . 22
13.2.9 Performance checks of dose and confidence limits estimation . 22
13.2.10 Performance checks for result report generation . 22
Annex A (informative) Sample data sheet for recording MN in BNCs .23
Annex B (informative) Instructions for requestor (sample) .24
Annex C (informative) Sample questionnaire .25
Annex D (informative) Sample of report for single assessment.27
Annex E (informative) Sample group report .29
Annex F (informative) Decision threshold and detection limit .31
Bibliography .34
iv
ISO 17099:2024(en)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards
bodies (ISO member bodies). The work of preparing International Standards is normally carried out through
ISO technical committees. Each member body interested in a subject for which a technical committee
has been established has the right to be represented on that committee. International organizations,
governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely
with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
The procedures used to develop this document and those intended for its further maintenance are described
in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the different types
of ISO document should be noted. This document was drafted in accordance with the editorial rules of the
ISO/IEC Directives, Part 2 (see www.iso.org/directives).
ISO draws attention to the possibility that the implementation of this document may involve the use of (a)
patent(s). ISO takes no position concerning the evidence, validity or applicability of any claimed patent
rights in respect thereof. As of the date of publication of this document, ISO had not received notice of (a)
patent(s) which may be required to implement this document. However, implementers are cautioned that
this may not represent the latest information, which may be obtained from the patent database available at
www.iso.org/patents. ISO shall not be held responsible for identifying any or all such patent rights.
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation of the voluntary nature of standards, the meaning of ISO specific terms and expressions
related to conformity assessment, as well as information about ISO's adherence to the World Trade
Organization (WTO) principles in the Technical Barriers to Trade (TBT), see www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 85, Nuclear energy, nuclear technologies, and
radiological protection, Subcommittee SC 2, Radiological protection, in collaboration with the European
Committee for Standardization (CEN) Technical Committee CEN/TC 430, Nuclear energy, nuclear technologies
and radiological protection, in accordance with the Agreement on technical cooperation between ISO and
CEN (Vienna Agreement).
This second edition cancels and replaces the first edition (ISO 17099:2014), which has been technically
revised.
The main changes are as follows:
— minor edits to text throughout;
— reorganization of document to better harmonize with other biological dosimetry standards;
— addition of 7.2.7 on data security plan;
— additional requirements added for the report on the conditions of the exposure for the calibration curve
in 10.2;
— relaxation of the number of individuals required for each age group for establishing background
micronucleus frequency, leaving the determination up to the head of the laboratory (10.3);
— addition of details on determining the minimal resolvable dose (10.4), the absorbed dose (11.2.4) and the
uncertainty (11.2.5);
— removal of reference to coefficient of variance when determining scoring expertise, focussing on the use
of 95 % confidence intervals to determine expertise (11.1.3);
— addition of reference to other exposure scenarios (11.2.8);
— removal of Annex on automated micronuclei scoring as it was deemed outside of the scope of the standard;
— addition of a sample group report (see Annex E);
v
ISO 17099:2024(en)
— addition of a detailed annex (see Annex F) for calculating the decision threshold and detection limit
along with a sample calculation and R script for performing these calculations.
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html.
vi
ISO 17099:2024(en)
Introduction
The purpose of this document is to define the use of the cytokinesis-block micronucleus (CBMN) assay with
human peripheral blood lymphocytes for biological dosimetry of exposure to ionizing radiation. This assay
is intended to be applied for accidental or malevolent exposures involving
a) up to a few casualties to provide individual whole-body dose estimates, or
b) in a triage mode to populations to provide rapid, lower accuracy dose estimates for individuals that can
be improved with more accurate analysis at a later time.
The CBMN assay is an alternative cytogenetic technique, which is possibly simpler and faster to perform
[1][2]
than the dicentric assay . It is also routinely used to demonstrate exposure to genotoxic agents, other
than ionizing radiation, which is not covered in this document. Although culture of the blood samples is
slightly longer than for dicentrics, the scoring of micronuclei (MN) in binucleated lymphocytes is easier.
As was done with the dicentric assay, the CBMN assay has been adapted for the emergency triage of large-
scale multi-casualty nuclear or radiological incident. The blood volume required for a sufficient number of
scorable binucleated cells (BNCs) is similar to that required for the dicentric assay. Again, the faster counting
speed for MN compensates for the extended culture time. However, it has to be considered that factors such
as age, sex, diet and environmental mutagens can have an influence on the results particularly after low
[3][4][5]
dose exposures . In addition, the CBMN assay can be performed in an automated mode using various
cytometric technologies but these are outside the scope of this document.
This document provides a guideline on how to perform the CBMN assay for dose assessment using
documented and validated procedures. Dose assessment using the CBMN assay has relevance in medical
management, radiation-protection management, record keeping, and medical/legal requirements. This
document is divided into two parts, according to the use of CBMN assay: radiation exposure of a few
individuals or population triage in a large radiological or nuclear event.
A part of the information in this document is contained in other international guidelines and scientific
publications, primarily in the International Atomic Energy Agency’s (IAEA) technical reports series on
biological dosimetry. However, this document expands and standardizes the quality assurance and quality
control, the criteria of accreditation and the evaluation of performance. This document is generally in
[6]
conformity with ISO/IEC 17025 with particular consideration given to the specific needs of biological
dosimetry. The expression of uncertainties in dose estimations given in this document complies with
[15] [7]
ISO/IEC Guide 98-3 (former GUM) and the ISO 5725 (all parts) .
vii
International Standard ISO 17099:2024(en)
Radiological protection — Performance criteria for
laboratories using the cytokinesis-block micronucleus
(CBMN) assay in peripheral blood lymphocytes for biological
dosimetry
1 Scope
This document gives guidance on
a) confidentiality of personal information for the customer and the laboratory,
b) laboratory safety requirements,
c) calibration sources and calibration dose ranges useful for establishing the reference dose-response
curves that contribute to the dose estimation from CBMN assay yields and the detection limit,
d) performance of blood collection, culturing, harvesting, and sample preparation for CBMN assay scoring,
e) scoring criteria,
f) conversion of micronucleus frequency in BNCs into an estimate of absorbed dose,
g) reporting of results,
h) quality assurance and quality control, and
i) informative annexes containing sample instructions for customers, sample questionnaire, a microscope
scoring data sheet, and a sample report.
This document excludes methods for automated scoring of CBMN.
2 Normative references
There are no normative references in this document.
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at https:// www .electropedia .org/
3.1
background frequency
background level
spontaneous yield (or number) of MN in BNCs recorded in control samples or individuals who are not
abnormally exposed to genotoxins including ionizing radiation
ISO 17099:2024(en)
3.2
binucleated cells
BNCs
cells that have completed one nuclear division after mitogen stimulation but have been blocked from
performing cytokinesis (3.6) and are the cell type in which micronuclei (3.9) are scored in the CBMN assay
Note 1 to entry: These cells are accumulated in culture using cytochalasin-B which is an inhibitor of cytokinesis.
3.3
chromosome
structure that comprises discrete packages of DNA and proteins that carries genetic information which
condense to form characteristically shaped bodies during nuclear division
3.4
confidence interval
statistical range about an estimated quantity within which the value of the quantity is expected to occur,
with a specified probability
3.5
cytochalasin-B
Cyto-B
reagent used to block cytokinesis (3.6) in dividing cells allowing once-divided cells to be identified as
binucleated cells
3.6
cytokinesis
physical process of cell division, which divides the cytoplasm of a parental cells into two daughter cells
3.7
dicentric
aberrant chromosome (3.3) bearing two centromeres derived from the joining of parts from two broken
chromosomes (3.3)
Note 1 to entry: It is generally accompanied by an acentric fragment.
3.8
linear energy transfer
LET
quotient of dE/dl, as defined by the International Commission on Radiation Units and Measurements (ICRU),
where dE is the average energy locally imparted to the medium by a charged particle of specific energy in
traversing a distance of dl
3.9
micronucleus
MN
small nucleus that arises from lagging acentric chromosome (3.3) fragments or whole chromosomes during
nuclear division and chromosome (3.3) segregation at mitosis during anaphase/telophase
Note 1 to entry: More that 90 % of the micronuclei induced by ionizing radiation arise from lagging acentric
chromosome fragments.
3.10
non-refractile
process by which materials do not have the ability to refract or scatter light
3.11
precision
dispersion of measurements with respect to a measure of location or central tendency
ISO 17099:2024(en)
3.12
quality assurance
planned and systematic actions necessary to provide adequate confidence that a process, measurement, or
service has satisfied given requirements for quality
3.13
quality control
part of quality assurance (3.12) intended to verify that systems and components correspond to pre-
determined requirements
3.14
service laboratory
laboratory performing biological dosimetry measurements
4 CMBN assay methodology used in this document
4.1 General
In this document, the frequency of MN in cultured human lymphocytes blocked in cytokinesis and scored by
microscopy is used for dose estimation after suspected exposure to ionizing radiation.
Lymphocytes are cultured by a method that permits once-divided cytokinesis-block cells to be recognized
by their binucleated appearance for analysis. This requires whole blood or isolated lymphocytes to be
incubated in culture medium with a mitogen that would enable scoring of MN in first-generation BNCs. A
cytokinesis blocking agent, cytochalasin-B, is added at least 6 h, i.e. approximately 24 h after the start of the
culture, before the first mitosis commences in order to arrest dividing lymphocytes at the BNC stage after
nuclear division is completed. The duration of the cell culture and the timing of addition of the arresting
agent are optimised to ensure an adequate frequency of binucleated cells.
BNCs are recovered from the cultures by centrifugation, placing in a hypotonic salt solution and fixing in
a mixture of methanol and acetic acid. Fixed cells are placed on microscope slides and stained. In the case
of isolated lymphocytes, it is also acceptable to prepare slides by cytocentrifugation of cells onto slides,
followed by air-drying, fixation with methanol, and staining. The exact protocol for cell culture, harvesting
BNCs and staining employed by a CBMN assay laboratory should be formally documented.
Microscope slides containing stained cells are scanned to identify suitable BNCs. The frequency of MN
observed in an appropriate number of scored BNCs is converted to an estimate of radiation dose by reference
to calibration data.
4.2 Requests for analysis and blood sampling
Depending on national regulations, the request for an analysis should normally be made by a physician
representing the patient, or the analysis could be requested by another authority due to legal claims. In
all cases where it is normally possible, the blood sampling for MN analysis shall be made with the patient’s
informed consent. The laboratory head, depending on the national regulations, may be required to maintain
the record of the patient's informed consent.
It is the responsibility of the medical staff (e.g. doctor, nurse, etc.) to schedule blood draw and shipping so as
to ensure that the blood sample is received by the laboratory in the best possible conditions (see 13.2.4). The
purpose is to avoid having the blood sample sit for several hours from time of blood draw and before sample
pickup for transportation (see Clause 5 for details).
5 Responsibility of the requestor
This clause includes items that are not controlled by the service laboratory. Prior to blood sampling, an
initial conversation between the requestor and the service laboratory should occur to co-ordinate the
sample collection and shipment. Specific requirements regarding sample collection and shipment should
be explained to the requestor including the approximate delivery time for the assay result(s). A standard
ISO 17099:2024(en)
instruction sheet (illustrated in Annex B) explaining the requirements should be sent to the requestor. The
requirements include:
1)
a) Blood sampling shall use Vacutainer® tubes containing lithium or sodium heparin as the anticoagulant
and the Vacutainer® tubes should either be sent or specified by the service laboratory.
b) Blood shall be collected (ideally about 5 ml), labelled accurately and unambiguously, maintained at
room temperature (around 20 °C), and sent to the service laboratory as soon as possible.
c) Precautions shall be taken to ensure the integrity of the container to prevent leakage during shipment.
Blood samples shall be kept at ambient temperature during shipment, i.e. 11° C to 30 °C. A temperature
recording device shall be included to ensure that the temperature during shipment is controlled.
Packaging and labelling shall conform to national and international regulations. If air transportation
is involved, a physical dosimeter shall be included to monitor whether the sample was irradiated in
transit.
d) A questionnaire (see Annex C) provided by the service laboratory shall be completed and returned prior
to the start of blood culturing.
e) The laboratory shall be alerted of biologically contaminated and/or infectious samples so that extra
precautions can be taken when handling the sample.
6 Responsibility of the service laboratory
6.1 Setup and sustainment of the quality assurance program
The service laboratory shall establish and maintain a quality assurance (QA) program (see Clause 13), which
covers all aspects of the service. The laboratory’s QA program shall address the following issues:
a) It shall include periodic internal checks of equipment operations, reagent suitability, and various
performance checks, i.e. intra-laboratory comparison exercises, operator qualifications, sample
protocol, scoring, dose estimations, report generation, etc.
b) It shall include periodic external checks of the laboratory’s operations. The external audits shall include
a review of the service laboratory’s documentation of equipment operations, reagent suitability, and
various performance checks, i.e. inter-comparison exercises, operator qualifications, sample transport
integrity, etc.
6.2 Responsibility during service
The service laboratory shall provide necessary guidance, procedures, and timely reporting to provide
dose assessment by the CBMN assay with a request for service. The service activities shall address the
following issues:
a) The service laboratory shall have documentation, reviewed and endorsed by a qualified expert,
i.e. service laboratory radiobiologist or equivalent, including the following:
1) an instruction sheet to be sent to the customer describing shipping procedures (see Annex B);
2) a questionnaire that shall elicit patient consent and all available information regarding the patient
and the exposure scenario (see Annex C);
3) step by step procedures for processing the blood sample from receipt of the sample to reporting of
the dose;
b) The service laboratory is not responsible for sample transport; however, they should provide advice
regarding sample transfer. If required, a kit for the collection of at least 5 ml whole blood in tubes
containing lithium or sodium heparin as the anticoagulant shall be sent to the requestor with the
V is an example of a suitable product available commercially. This information is given for the convenience of users of this document and does
1) acutainer
not constitute an endorsement by ISO of this product Equivalent products may be used if they can be shown to lead to the same results.
.
ISO 17099:2024(en)
appropriately labelled and addressed packaging material for the return of the sample to the service
laboratory. The packaging shall conform to national and/or international regulations for the transit of
potentially infectious pathological specimens (see 13.2.4).
c) After receipt of the blood sample, the following steps shall be performed:
1) document the receipt of the blood sample (date, time, recipient);
2) check for conformity of the sample (blood volume, integrity of the tubes);
3) mark the blood sample with a unique code;
4) store samples at room temperature and document the place of storage until the setting up of
cultures;
5) set up cultures in parallel as soon as possible and document date, time, and operator;
6) document all the reagents used for culturing with appropriate lot numbers and expiry dates;
7) document the addition of reagents and the end of the culture (date, time, operator);
8) document the short- and long-term storage of the sample until slide making;
9) document the slide codes, number of slides, and location of storage;
10) document the results from scoring;
11) store the slides and case documents in an appropriate place for possible medico-legal re-evaluation
of the case;
d) The service laboratory shall interpret the results and prepare reports (see Annex D).
e) The service laboratory shall sustain a dialogue with and provide results to the requestor.
7 Confidentiality of personal information
7.1 Overview
Biological dosimetry investigations made by a service laboratory shall be undertaken in accordance
with national regulations regarding confidentiality. This would normally include the maintenance of
confidentiality of all of the patient's information including identity, medical data, etc. In addition, the
commercial confidentiality of the patient's employer and any other organizations involved in a radiological
or nuclear accident/incident should be observed.
This requirement extends to
a) written, electronic, or verbal communications between the laboratory and the person/organization
requesting the analysis and receiving the report, and
b) the secure protection of confidential information held within the organization where the service
laboratory is located.
7.2 Applications of the principle of confidentiality
7.2.1 Delegation of responsi
...
Frequently Asked Questions
SIST EN ISO 17099:2024 is a standard published by the Slovenian Institute for Standardization (SIST). Its full title is "Radiological protection - Performance criteria for laboratories using the cytokinesis block micronucleus (CBMN) assay in peripheral blood lymphocytes for biological dosimetry (ISO 17099:2024)". This standard covers: This document gives guidance on a) confidentiality of personal information for the customer and the laboratory, b) laboratory safety requirements, c) calibration sources and calibration dose ranges useful for establishing the reference dose-response curves that contribute to the dose estimation from CBMN assay yields and the detection limit, d) performance of blood collection, culturing, harvesting, and sample preparation for CBMN assay scoring, e) scoring criteria, f) conversion of micronucleus frequency in BNCs into an estimate of absorbed dose, g) reporting of results, h) quality assurance and quality control, and i) informative annexes containing sample instructions for customers, sample questionnaire, a microscope scoring data sheet, and a sample report. This document excludes methods for automated scoring of CBMN.
This document gives guidance on a) confidentiality of personal information for the customer and the laboratory, b) laboratory safety requirements, c) calibration sources and calibration dose ranges useful for establishing the reference dose-response curves that contribute to the dose estimation from CBMN assay yields and the detection limit, d) performance of blood collection, culturing, harvesting, and sample preparation for CBMN assay scoring, e) scoring criteria, f) conversion of micronucleus frequency in BNCs into an estimate of absorbed dose, g) reporting of results, h) quality assurance and quality control, and i) informative annexes containing sample instructions for customers, sample questionnaire, a microscope scoring data sheet, and a sample report. This document excludes methods for automated scoring of CBMN.
SIST EN ISO 17099:2024 is classified under the following ICS (International Classification for Standards) categories: 13.280 - Radiation protection; 71.040.10 - Chemical laboratories. Laboratory equipment. The ICS classification helps identify the subject area and facilitates finding related standards.
SIST EN ISO 17099:2024 has the following relationships with other standards: It is inter standard links to SIST EN ISO 17099:2017. Understanding these relationships helps ensure you are using the most current and applicable version of the standard.
You can purchase SIST EN ISO 17099:2024 directly from iTeh Standards. The document is available in PDF format and is delivered instantly after payment. Add the standard to your cart and complete the secure checkout process. iTeh Standards is an authorized distributor of SIST standards.
Die SIST EN ISO 17099:2024 ist ein bedeutendes Dokument, das sich mit den Leistungsanforderungen für Labore befasst, die den Zyto-Kinese-Block-Mikronukleus (CBMN) Test zur biologischen Dosimetrie in peripheren Blutlymphozyten einsetzen. Diese Norm deckt einen umfassenden Bereich ab, der nicht nur professionelle Standards definiert, sondern auch die Sicherheit und Vertraulichkeit im Laboralltag gewährleistet. Ein herausragender Aspekt der Norm ist die detaillierte guidance zur Vertraulichkeit persönlicher Informationen, was besonders in der heutigen Zeit von großer Relevanz ist. Dieses Element fördert das Vertrauen zwischen Kunden und Laboren und verbessert somit die Gesamtverfahrensweise. Des Weiteren legt die SIST EN ISO 17099:2024 klar definierte Sicherheitsanforderungen fest, die unerlässlich sind, um ein sicheres Arbeitsumfeld zu gewährleisten. Dies ist nicht nur vorteilhaft für die Laborangestellten, sondern auch für die Qualität der durchgeführten Tests. Ein weiterer Stärke dieser Norm ist die umfassende Anleitung zu Kalibrierungsquellen und Kalibrierungsdosisbereichen, die für die Etablierung der Referenz-Dosis-Wirkungs-Kurven wichtig sind. Diese Informationen sind entscheidend für die genaue Schätzung der Dosis, die aus den Ergebnissen des CBMN-Tests abgeleitet wird. Die Norm behandelt auch die Durchführung von Blutabnahme, Kultur, Ernte und Probenvorbereitung für die Bewertung des CBMN-Tests. Die klaren Richtlinien zu diesen Prozessen verbessern die Konsistenz und Zuverlässigkeit der Testergebnisse. Darüber hinaus werden spezifische Bewertungskriterien und die Umwandlung der Mikronukleusfrequenz in absorbierte Dosen thematisiert, was für die genaue Datenauswertung unerlässlich ist. Besonders hervorzuheben ist der Abschnitt über die Ergebnisberichterstattung, der sicherstellt, dass die Informationen klar und präzise vermittelt werden. Darüber hinaus bietet die Norm umfassende Richtlinien für Qualitätsmanagement und -kontrolle, die für jedes Labor von zentraler Bedeutung sind. Informative Anhänge, die Musteranweisungen für Kunden, einen Fragebogen, ein Mikroskopbewertungsdatenblatt und einen Musterbericht enthalten, runden das Dokument ab und machen es zu einem wertvollen Hilfsmittel für die Praxis. Zusammenfassend lässt sich sagen, dass die SIST EN ISO 17099:2024 eine gut strukturierte und relevante Norm für Labore ist, die den CBMN-Test durchführen, und einen wesentlichen Beitrag zur biologischen Dosimetrie leistet.
SIST EN ISO 17099:2024は、個人情報の機密性、実験室の安全要件、及びCBMNアッセイのための参照線量-応答曲線を確立するために有用な校正源と校正線量範囲の設定に関するガイダンスを提供します。この標準のスコープは広範であり、血液サンプルの採取、培養、収穫、及びCBMNアッセイのスコアリングに必要なサンプル準備におけるパフォーマンスを促進します。 本標準は、CBMNアッセイによる生物 dosimetryにおけるパフォーマンス基準を定義しており、スコアリング基準の明確な指針を提供します。これにより、微小核の頻度をBNCから吸収線量の推定値に変換する手法も明記されています。また、結果の報告手順や、品質保証と品質管理に関する指針も含まれており、全体的な実験室の運用の質を向上させることができます。 さらに、顧客向けのサンプル指示、質問票、顕微鏡スコアリングデータシート、及びサンプル報告書を含む情報的付録が添付されており、利用者が容易にこの標準を適用できるように工夫されています。これにより、研究者や技術者がCBMNアッセイを用いて正確な生物 dosimetryを行うための基盤が整備されています。 SIST EN ISO 17099:2024は、実験室における安全性と精度の確保を重視しており、その適用は特に放射線の影響を評価するための重要なステップとなります。この標準は、医療や環境モニタリングの分野においても極めて関連性が高く、実験の信頼性確保に寄与するため、研究者や施設にとって不可欠な文書です。
Le document SIST EN ISO 17099:2024 présente des directives cruciales pour la protection radiologique, en particulier concernant le critère de performance des laboratoires utilisant le test de micronoyaux par bloc de cytokines (CBMN) sur des lymphocytes de sang périphérique pour la dosimétrie biologique. Cette norme se distingue par son ample portée, abordant des aspects essentiels pour assurer la fiabilité et la qualité des analyses effectuées en laboratoire. Parmi ses points forts, le document s'attarde sur la confidentialité des informations personnelles tant pour le client que pour le laboratoire, garantissant ainsi une protection adéquate des données sensibles. Les exigences de sécurité en laboratoire sont également mises en avant, ce qui est primordial pour maintenir un environnement de travail sécurisé pour le personnel et les échantillons. La normalisation des sources de calibration et des plages de doses de calibration est un aspect essentiel, car elle permet d’établir des courbes dose-réponse de référence cruciales pour l'estimation de dose à partir des rendements du test CBMN. De plus, le document offre des lignes directrices détaillées sur la collecte de sang, la culture, la récolte et la préparation des échantillons en vue du scoring du CBMN, contribuant ainsi à une approche standardisée et fiable. Les critères de scoring clairement définis ainsi que la conversion de la fréquence des micronoyaux en BNC en une estimation de dose absorbée garantissent la cohérence dans l’interprétation des résultats. La normée stipule également la nécessité d'une communication claire des résultats, renforçant la transparence et la compréhension entre le laboratoire et ses clients. Enfin, la norme aborde la qualité assurance et le contrôle de qualité, des éléments fondamentaux pour la crédibilité des résultats. Les annexes informatives fournissent des instructions pratiques, un questionnaire type, une fiche de données pour le scoring au microscope et un rapport type, offrant ainsi aux utilisateurs des outils utiles pour l’application de la norme. En somme, la SIST EN ISO 17099:2024 constitue une référence indispensable pour les laboratoires engagés dans la dosimétrie biologique, apportant une structure robuste pour garantir des pratiques de protection radiologique efficaces et fiables.
SIST EN ISO 17099:2024는 방사선 보호와 관련된 실험실에서 말초 혈림프구의 세포 분열 차단 미세핵(CBMN) 검사를 사용한 생물학적 선량 측정에 대한 성능 기준을 제시하는 중요한 표준입니다. 이 문서의 범위는 개인 정보의 기밀성, 실험실 안전 요구 사항, 그리고 CBMN 검사로부터의 선량 추정에 기여하는 기준선 반응 곡선을 설정하는 데 유용한 교정소스 및 교정 선량 범위와 같은 다양한 요소를 다룹니다. 이 표준의 주요 강점은 실험실의 혈액 수집, 배양, 수확 및 CBMN 검사 채점에 대한 성능 기준을 명확히 제시함으로써 실험의 신뢰성을 향상시키는 데 기여한다는 점입니다. 또한 미세핵 빈도를 BNC로의 변환을 통해 흡수 선량을 추정하는 방법을 표기함으로써 과학적 근거를 정립하고 있습니다. 결과 보고 및 품질 보증, 품질 관리에 대한 지침이 포함되어 있어 실험실의 운영 전반에 걸쳐 신뢰성을 높이는 데 중요한 역할을 합니다. SIST EN ISO 17099:2024는 또한 고객을 위한 샘플 지침, 설문지, 현미경 채점 데이터 시트 및 샘플 보고서와 같은 정보 부록을 포함하고 있어 실험실이 데이터를 효과적으로 관리하고 보고할 수 있는 방법을 제공합니다. 이는 실제 실험실 운영과 관련하여 매우 중요한 요소입니다. 이러한 모든 요소는 CBMN 검사를 이용한 생물학적 선량 측정의 유효성을 높이고, 안전성을 강화하는 동시에 신뢰할 수 있는 데이터를 제공함으로써 방사선 보호 분야에서 이 표준의 관련성을 더욱 강조합니다.
The SIST EN ISO 17099:2024 standard is a crucial document that outlines performance criteria for laboratories employing the cytokinesis block micronucleus (CBMN) assay in peripheral blood lymphocytes for biological dosimetry. Its comprehensive scope addresses multiple key elements necessary for the reliable and ethical conduct of biological dosimetry, emphasizing the importance of radiological protection. One of the standard's significant strengths is its focus on confidentiality, ensuring that personal information of customers and laboratory staff is adequately protected. This inclusion is particularly vital in the context of biological dosimetry, where sensitive health data might be involved. Furthermore, the document establishes rigorous laboratory safety requirements, promoting a secure environment for both personnel and samples involved in the CBMN assay. Additionally, the standard provides clear guidelines on calibration sources and dose ranges, which are essential for developing accurate reference dose-response curves. These curves directly affect dose estimations derived from CBMN assay yields and the detection limits, establishing a robust foundation for radiological assessments. This precision is enhanced by specific criteria for blood collection, culturing, harvesting, and sample preparation, ensuring that each step in the CBMN assay scoring process meets high standards of quality and reliability. The scoring criteria outlined in SIST EN ISO 17099:2024 further enhance its utility, guiding laboratories in the effective interpretation of micronucleus frequency in binucleated cells (BNCs) as it pertains to absorbed dose estimations. Reporting of results is another critical aspect covered, providing a clear framework for communication of findings in a format that is understandable to stakeholders. Quality assurance and quality control measures detailed in the standard ensure that laboratories maintain high operational standards, reinforcing the credibility of biological dosimetry outcomes derived from CBMN assays. The inclusion of informative annexes, such as sample instructions for customers, a sample questionnaire, a microscope scoring data sheet, and an example report, adds practical value, facilitating ease of implementation and adherence to the standard. SIST EN ISO 17099:2024 effectively excludes methods for automated scoring of the CBMN, which emphasizes its focus on maintaining a consistent manual approach that ensures accuracy and reliability in results. Overall, this standard plays an essential role in establishing best practices in radiological protection for laboratories utilizing the CBMN assay, making it an indispensable reference for professionals in the field of biological dosimetry.
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