prEN 18000-3

Animal health diagnostic analyses - Control of in vitro diagnostic reagents - Part 3: Reagents for PCR techniques

This document specifies the control and approval of in vitro diagnostic reagents used in animal health for the detection, and/or absolute quantification of pathogen-specific nucleic acid (DNA or RNA) by PCR (e.g. endpoint PCR, real-time PCR, reverse transcription-PCR).
This document is applicable to diagnostic reagents as a priority for infectious diseases (due to bacteria, viruses, fungi, or parasites, including genetic markers associated with pathogenicity, such as antimicrobial resistance or toxin production) and associated animal species for which harmonization of practices in this area is needed, i.e. those for which the national, regional or international regulatory framework provides for the control of trade in animals and/or animal products and/or the definition of a health status (absence of infection) of areas, establishments or individuals. Anyhow, all reagents designated by the competent authorities fall under the scope of this document. Nevertheless, the authorities or any other animal health stakeholder can choose to derogate in specific and very limited situations such as emerging, exotic or rare diseases.
This document is not applicable to all existing diagnostic reagents, in particular those for which certain parameters described in this document cannot be validly evaluated in accordance with international requirements, due, e.g. to the absence of a specific reference standard and/or accessible and duly validated reference materials.
The PCR diagnosis usually involves the use of a nucleic acid extraction and/or purification reagent, and a PCR reagent. The PCR method (when applicable) involves the successive use of these distinct reagents. PCR reagent control can be performed if the applicant provides evidence of the validity of the PCR reagent for use in the animal health diagnostic analysis, by proving its diagnostic performances with nucleic acid extracts obtained from the different matrices described in the instruction for use. The control of a complete PCR method by the applicant and the control organization is performed only if the PCR reagent cannot be dissociated from an nucleic acid extraction and/or purification systems. This document does not cover the control of the nucleic acid extraction and/or purification reagents, only.
This document does not cover the step in which the user verifies a reagent (analysis method adoption).
NOTE   Prion diseases are not included in the scope of this third part of the EN 18000 series. Unlike other infectious diseases, prion diseases are not diagnosed using PCR assays because prions lack a nucleic acid component and consist solely of an abnormally folded conformer of the normal host protein.

Tiergesundheitsdiagnostische Analysen - Kontrolle von in-vitro-diagnostischen Reagenzien - Teil 3: Reagenzien für PCR Verfahren

Dieses Dokument legt die Prüfung und Zulassung von im Bereich der Tiergesundheit verwendeten in-vitro-diagnostischen Reagenzien zum Nachweis und/oder zur absoluten Quantifizierung von erregerspezifischer Nukleinsäure (DNA oder RNA) durch PCR (z. B. Endpunkt-PCR, Real-Time-PCR, Reverse-Transkription-PCR) fest.
Dieses Dokument gilt für diagnostische Reagenzien, vorrangig für Infektionskrankheiten (durch Bakterien, Viren, Pilze oder Parasiten, einschließlich mit Pathogenität verbundener genetischer Marker, beispielsweise für antimikrobielle Resistenz oder Toxinproduktion) und entsprechende Tierarten, für die eine Harmonisierung der Praktiken in diesem Bereich erforderlich ist, d. h. für diejenigen, für die der nationale, regionale oder internationale Regelungsrahmen die Kontrolle des Handels mit Tieren und/oder tierischen Erzeugnissen und/oder die Festlegung eines Gesundheitsstatus (Infektionsfreiheit) von Gebieten, Einrichtungen oder Personen vorsieht. Gleichwohl fallen alle von den zuständigen Behörden festgelegten Reagenzien in den Anwendungsbereich dieses Dokuments. Nichtsdestotrotz können die Behörden oder andere interessierte Parteien im Bereich der Tiergesundheit sich dazu entschließen, in bestimmten und eng umrissenen Situationen, beispielsweise bei neu aufkommenden, exotischen oder seltenen Krankheiten, davon abzuweichen.
Dieses Dokument ist nicht auf alle vorhandenen diagnostischen Reagenzien anwendbar, insbesondere nicht auf solche, für die bestimmte in diesem Dokument beschriebene Parameter nicht in Übereinstimmung mit internationalen Anforderungen gültig bewertet werden können, z. B. weil kein spezifisches Referenznormal und/oder keine zugänglichen und ordnungsgemäß validierten Referenzmaterialien zur Verfügung stehen.
Die PCR-Diagnostik umfasst üblicherweise die Anwendung einer Nukleinsäure-Extraktion und/oder eines Reinigungsreagenzes und eines PCR-Reagenzes. Das PCR-Verfahren (sofern anwendbar) umfasst die aufeinanderfolgende Verwendung dieser verschiedenen Reagenzien. Die Prüfung des PCR-Reagenzes kann durchgeführt werden, wenn der Antragsteller einen Nachweis der Validität des PCR-Reagenzes für die Verwendung in diagnostischen Untersuchungen im Bereich der Tiergesundheit vorlegt, indem seine diagnostischen Leistungsmerkmale bei Verwendung mit Nukleinsäure-Extrakten, die aus den in der Gebrauchsinformation beschriebenen verschiedenen Matrices erhalten wurden, nachgewiesen werden. Die Prüfung eines vollständigen PCR-Verfahrens durch den Antragsteller und die Prüfstelle wird nur durchgeführt, wenn das PCR-Reagenz nicht von Nukleinsäure-Extraktions- und/oder -Reinigungssystemen gesondert geprüft werden kann. Die alleinige Prüfung der Reagenzien für Nukleinsäure-Extraktion und/oder -Reinigung wird in diesem Dokument nicht behandelt.
In diesem Dokument wird nicht der Schritt der Verifizierung eines Reagenzes durch den Anwender behandelt (Annahme eines Analyseverfahrens).
ANMERKUNG   Durch Prionen verursachte Krankheiten sind nicht im Anwendungsbereich dieses dritten Teils der Normenreihe EN 18000 enthalten. Im Unterschied zu anderen Infektionskrankheiten werden durch Prionen verursachte Krankheiten nicht mithilfe von PCR-Tests diagnostiziert, weil Prionen ein Nukleinsäure-Bestandteil fehlt und sie lediglich aus einem anormal gefalteten Konformer des normalen Wirtsproteins bestehen.

Analyses de diagnostic en santé animale - Contrôle des réactifs de diagnostic in vitro - Partie 3 : Réactifs pour les techniques PCR

Le présent document spécifie le contrôle et l’approbation des réactifs de diagnostic in vitro utilisés en santé animale pour la détection, et/ou la quantification absolue d’acide nucléique spécifique à un pathogène (ADN ou ARN) par PCR (par exemple, PCR en point final, PCR en temps réel, PCR à transcription inverse).
Le présent document s’applique aux réactifs de diagnostic, en priorité pour les maladies infectieuses (dues à des bactéries, des virus, des champignons ou des parasites, y compris des marqueurs génétiques associés à une pathogénicité telle qu’une résistance antimicrobienne ou la production de toxines) et les espèces animales associées pour lesquelles une harmonisation des pratiques dans ce domaine est nécessaire, c’est-à-dire celles pour lesquelles le cadre réglementaire national, régional ou international prévoit le contrôle des échanges d’animaux et/ou de produits animaux et/ou la définition d’un état sanitaire (absence d’infection) des zones, établissements ou individus. Quoi qu’il en soit, tous les réactifs désignés par les autorités compétentes relèvent du domaine d’application du présent document. Néanmoins, les autorités ou toute autre partie prenante de la santé animale peuvent choisir de déroger à ces exigences dans des situations spécifiques et très limitées, telles que des maladies émergentes, exotiques ou rares.
Le présent document ne s’applique pas à tous les réactifs de diagnostic existants, en particulier ceux pour lesquels certains paramètres décrits dans le présent document ne peuvent pas être évalués de manière valable conformément aux exigences internationales en raison, par exemple, de l’absence d’une norme de référence spécifique et/ou de matériaux de référence accessibles et dûment validés.
Le diagnostic par PCR implique généralement l’utilisation d’un réactif d’extraction de l’acide nucléique et/ou de purification de l’acide nucléique, et d’un réactif de PCR. La méthode de PCR (lorsqu’elle est applicable) implique l’utilisation successive de ces différents réactifs. Le contrôle des réactifs de PCR peut être réalisé si le demandeur fournit la preuve de la validité du réactif de PCR pour utilisation dans le cadre de l’analyse diagnostique en santé animale, en mettant à disposition ses performances de diagnostic avec les extraits d’acide nucléique obtenus à partir des différentes matrices décrites dans la notice d’utilisation. Le contrôle d’une méthode de PCR complète par le demandeur et l’organisme de contrôle est réalisé uniquement si le réactif de PCR est indissociable des systèmes d’extraction de l’acide nucléique et/ou de purification de l’acide nucléique. Le présent document ne couvre pas le contrôle des réactifs d’extraction de l’acide nucléique et/ou de purification de l’acide nucléique uniquement.
Le présent document ne couvre pas l’étape de vérification d’un réactif par l’utilisateur (adoption de la méthode d’analyse).
NOTE   Les maladies à prions ne relèvent pas du domaine d’application de la présente troisième partie de la série EN 18000. Contrairement à d’autres maladies infectieuses, les maladies à prions ne sont pas diagnostiquées à l’aide d’analyses par PCR, car les prions ne contiennent pas d’acide nucléique et se composent uniquement d’un conformère anormalement replié de la protéine hôte normale.

Diagnostične analize zdravja živali - Nadzor diagnostičnih reagentov in vitro - 3. del: Reagenti za metode PCR

General Information

• Status: Not Published
• Publication Date: 24-Nov-2026
• ICS: 11.220 - Veterinary medicine
• Technical Committee: CEN/TC 469 - Animal Health
• Drafting Committee: CEN/TC 469/WG 1 - Reagents and methods
• Current Stage: 4020 - Submission to enquiry - Enquiry
• Start Date: 03-Apr-2025
• Due Date: 09-Jul-2025
• Completion Date: 03-Apr-2025
• Ref Project: oSIST prEN 18000-3:2025 - Animal health diagnostic analyses - Control of in vitro diagnostic reagents - Part 3: Reagents for PCR techniques

Standards Content (Sample)

SLOVENSKI STANDARD
01-junij-2025
Diagnostične analize zdravja živali - Nadzor diagnostičnih reagentov in vitro - 3.
del: Reagenti za metode PCR
Animal health diagnostic analyses - Control of in vitro diagnostic reagents - Part 3:
Reagents for PCR techniques
Tiergesundheitsdiagnostische Analysen - Kontrolle von in-vitro-diagnostischen
Reagenzien - Teil 3: Reagenzien für PCR Verfahren
Analyses de diagnostic en santé animale - Contrôle des réactifs de diagnostic in vitro -
Partie 3 : Réactifs pour les techniques PCR
Ta slovenski standard je istoveten z: prEN 18000-3
ICS:
11.100.10 Diagnostični preskusni In vitro diagnostic test
sistemi in vitro systems
11.220 Veterinarstvo Veterinary medicine
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

DRAFT
EUROPEAN STANDARD
NORME EUROPÉENNE
EUROPÄISCHE NORM
April 2025
ICS 11.220
English Version
Animal health diagnostic analyses - Control of in vitro
diagnostic reagents - Part 3: Reagents for PCR techniques
Analyses de diagnostic en santé animale - Contrôle des Tiergesundheitsdiagnostische Analysen - Kontrolle von
réactifs de diagnostic in vitro - Partie 3 : Réactifs pour in-vitro-diagnostischen Reagenzien - Teil 3:
les techniques PCR Reagenzien für PCR Verfahren
This draft European Standard is submitted to CEN members for enquiry. It has been drawn up by the Technical Committee
CEN/TC 469.
If this draft becomes a European Standard, 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.

This draft European Standard was established by CEN 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.
Recipients of this draft are invited to submit, with their comments, notification of any relevant patent rights of which they are
aware and to provide supporting documentation.

Warning : This document is not a European Standard. It is distributed for review and comments. It is subject to change without
notice and shall not be referred to as a European Standard.

EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2025 CEN All rights of exploitation in any form and by any means reserved Ref. No. prEN 18000-3:2025 E
worldwide for CEN national Members.

Contents Page
European foreword . 3
Introduction . 4
1 Scope . 5
2 Normative references . 5
3 Terms and definitions . 5
4 General control steps . 10
5 Prerequisites of the PCR reagent control for the control organisation . 10
5.1 General . 10
5.2 Reference material . 10
5.3 Définition of the purpose of the PCR reagents and of the PCR method when applicable . 11
5.4 Additional useful information . 11
6 Initial conformity control . 11
6.1 General . 11
6.2 Characterization of the reagents by the applicant and documentary review by the
control organization . 11
6.2.1 General . 11
6.2.2 Définition of the interpretation method and threshold(s) . 14
6.2.3 Analytical spécificity of the PCR reagent . 14
6.2.4 Analytical sensitivity of the PCR reagent (LOD and MDL ) . 15
PCR PCR
6.2.5 Operating range of the quantitative PCR reagent (linearity, amplification éfficiéncy and
limit of quantification) when applicable . 15
6.2.6 Within-laboratory reproducibility of the PCR reagent . 16
6.2.7 Analytical sensitivity of the PCR method (MDL ) when applicable . 16
METHOD
6.2.8 Accuracy of the quantitative PCR method (operating range and limits of quantification)
when applicable . 16
6.2.9 Within-laboratory reproducibility of the PCR method when applicable . 17
6.2.10 Interlaboratory reproducibility of the PCR method when applicable . 17
6.2.11 Diagnostic sensitivity and diagnostic spécificity . 17
6.2.12 Validation of the conditions of use - Robustness . 17
6.2.13 Vérification of the stability . 17
6.3 Initial control of the reagents by the control organisation . 18
6.3.1 General . 18
6.3.2 Analytical spécificity of the PCR reagent . 18
6.3.3 Analytical sensitivity of the PCR reagent (MDL ) . 19
PCR
6.3.4 Operating range of the quantitative real-time PCR reagent (linearity, amplification
éfficiéncy and limit of quantification) . 19
6.3.5 Analytical sensitivity of the PCR method (MDL ), when applicable . 19
METHOD
6.3.6 Diagnostic sensitivity and spécificity of the PCR method when applicable . 19
6.3.7 Accuracy of the quantitative PCR method (validity range and limit of quantification),
when applicable . 19
7 Batch-to-batch control . 19
7.1 Control at the start of the batch shelf-life . 19
7.2 Control during the batch shelf-life . 20
7.3 Derogations from systematic batch-to-batch control . 20
8 Special cases . 20
8.1 Multiple protocols . 20
8.2 Multiple matrices . 20
8.3 Pooling of samples . 20
Bibliography . 22
European foreword
This document (prEN 18000-3:2025) has been prepared by Technical Committee CEN/TC "Animal
health diagnostic analyses", the secretariat of which is held by AFNOR.
This document is currently submitted to the CEN Enquiry.
Introduction
The purpose of the EN 18000 series is to facilitate the mutual recognition of the work of the animal
health in vitro diagnostic reagent control organisations at European level (or even more widely) and
thus to eventually allow the use of strategic reagents controlled by a single third-party control
organization for a given disease.
The EN 18000 series establishes the requirements for the control of in vitro diagnostic reagents in animal
health. This series is divided into three parts.
The first part concerns terms and définitions, and the submission of a reagent dossier to a control
organization for control and approval.
The second part concerns the spécific aspects of the control by such organizations of an immunological
diagnostic reagent.
The third part concerns the spécific aspects of the control by such organizations of a polymerase-chain
reaction (PCR) diagnostic reagent for the detection or quantification of pathogén-spécific nucleic acid. It
involves control organizations (CO) and applicants (including their subcontractors, when relevant).
Like any standard, this document is intended to be voluntary and, if its use is prescribed by a competent
authority or any other animal health stakeholder, it will be up to them to determine for which diseases
and to which extent this document will be applied by the control bodies they have designated for this
purpose.
1 Scope
This document spécifiés the control and approval of in vitro diagnostic reagents used in animal health
for the detection, and/or absolute quantification of pathogén-spécific nucleic acid (DNA or RNA) by PCR
(e.g. endpoint PCR, real-time PCR, reverse transcription-PCR).
This document is applicable to diagnostic reagents as a priority for infectious diseases (due to bacteria,
viruses, fungi, or parasites, including genetic markers associated with pathogenicity, such as
antimicrobial resistance or toxin production) and associated animal species for which harmonization
of practices in this area is needed, i.e. those for which the national, regional or international regulatory
framework provides for the control of trade in animals and/or animal products and/or the définition
of a health status (absence of infection) of areas, establishments or individuals. Anyhow, all reagents
designated by the competent authorities fall under the scope of this document. Nevertheless, the
authorities or any other animal health stakeholder can choose to derogate in spécific and very limited
situations such as emerging, exotic or rare diseases.
This document is not applicable to all existing diagnostic reagents, in particular those for which certain
parameters described in this document cannot be validly evaluated in accordance with international
requirements, due, e.g. to the absence of a spécific reference standard and/or accessible and duly
validated reference materials.
The PCR diagnosis usually involves the use of a nucleic acid extraction and/or purification reagent, and
a PCR reagent. The PCR method (when applicable) involves the successive use of these distinct
reagents. PCR reagent control can be performed if the applicant provides evidence of the validity of the
PCR reagent for use in the animal health diagnostic analysis, by proving its diagnostic performances
with nucleic acid extracts obtained from the different matrices described in the instruction for use. The
control of a complete PCR method by the applicant and the control organization is performed only if the
PCR reagent cannot be dissociated from an nucleic acid extraction and/or purification systems. This
document does not cover the control of the nucleic acid extraction and/or purification reagents, only.
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