ISO INTERNATIONAL STANDARD 16140-2 First edition 2016-06-15 Microbiology of the food chain - Method validation - Part 2: Protocol for the validation of alternative (proprietary) methods against a reference method Microbiologie de la chaine alimentaire - Validation des methodes - Partie 2: Protocole pour la validation de méthodes alternatives (commerciales) par rapport a une méthode de réference Reference number IS0 16140-2:2016(E) ISO International Organization for Standardization @ IS0 2016 HEJIANG INST OF STANDARDIZATION C1 5956617 ed without license from IHS IS0 16140-2:2016(E) COPYRIGHT PROTECTEDDOCUMENT IS0 2016, Published in Switzerland All rights reserved. Unless otherwise specified, 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 the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address below or ISO's member body in the country of the requester. ISO copyright office Ch. de Blandonnet 8 . CP 401 CH-1214 Vernier, Geneva, Switzerland Tel. +41 22 749 01 11 Fax +41 22 749 09 47 [email protected] www.iso.org Licensee-ZHEJIANG INST OF STANDARDIzRoISQ 6 - All rights reserved Not for Resale, 2016/8/16 08:40:13 networking permited without license from IHS IS0 16140-2:2016(E) Contents Page Foreword ..iv Introduction 1 Scope. .1 2 Normative references .1 3 Terms and definitions 4 General principles for the validation of alternative methods 1 5 Qualitative methods - Technical protocol for validation 2 5.1 Method comparison study .2 5.1.1 General considerations. 2 5.1.2 Paired or unpaired study .2 5.1.3 Sensitivity study . 2 5.1.4 Relative level of detection study .7 5.1.5 Inclusivity and exclusivity study ..9 5.2 Interlaboratory study 10 5.2.1 General considerations .10 5.2.2 Measurement protocol .10 5.2.3 Calculations and summary of data ..12 5.2.4 Interpretation of data 15 6 Quantitative methods - Technical protocol for validation ..16 6.1 Method comparison study ..16 6.1.1 General considerations .16 6.1.2 Relative trueness study .16 6.1.3 Accuracyprofilestudy 20 6.1.4 Limit of quantification study 24 6.1.5 Inclusivity and exclusivity study 24 6.2 Interlaboratory study 26 6.2.1 General considerations. .26 6.2.2 Measurement protocol .26 6.2.3 Calculations, summary, and interpretation of data. .27 Annex A (informative) Classification of sample types and suggested target combinations for validation studies .30 Annex B (normative) Order of preference for use of naturally and artificially contaminated samples in validation studies. .46 Annex C (informative) General protocols for contamination by mixture and artificial contaminationoffoods ..47 Annex D (informative) Models for RLoD calculations using data from the method comparison study .50 Annex E (normative) Points to be considered when selecting strains for testing inclusivity and exclusivity .52 Annex F (informative) Considerations for calculations of the relative level of detection (RLOD)betweenlaboratoriesasobtainedinaninterlaboratorystudy .54 Annex G (informative) Principle of the accuracy profile for validation of quantitative models ....57 Annex H (informative) Application of the accuracy profile in the method comparison study...59 Annex I (informative) Example of the application of the accuracy profile for an interlaboratory study .62 Bibliography .66 iii vithout license from IHS IS0 16140-2:2016(E) 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