ISO INTERNATIONAL STANDARD 20813 First edition 2019-05 Molecular biomarker analysis Methods of analysis for the detection and identification of animal species in foods and food products (nucleic acid-based methods) General requirements and definitions Analyse moléculaire de biomarqueurs - Méthodes d'analyse pour la détection et I'identification des especes animales dans les aliments et les produits alimentaires (méthodes basées sur I'utilisation des acides nucleiques) Exigences générales et définitions Reference number IS0 20813:2019(E) os1 @ IS0 2019 IS0 20813:2019(E) COPYRIGHTPROTECTEDDOCUMENT @ ISO 2019 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 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 CP 401 : Ch. de Blandonnet 8 CH-1214 Vernier, Geneva Phone: +4122 749 0111 Fax: +41 22 749 09 47 Email: [email protected] Website: www.iso.org Published in Switzerland ii @ IS0 2019 - All rights reserved IS0 20813:2019(E) Contents Page Foreword ..iv 1 Scope .1 2 Normative references ..1 3 Terms and definitions 4 Performance characteristics of the methods .2 4.1 General 2 4.2 Scope of the method. .2 4.3 Scientific basis 22 4.4 Units of measurement 4.5 Applicability 2 4.6 Specificity .3 4.6.1 General 3 4.6.2 Requirements for inclusivity testing 3 4.6.3 Requirements for exclusivity testing .3 4.7 Sensitivity 4 4.7.1 General .4 4.7.2 Limit of detection (LOD) 4 4.8 Specific requirements for quantitative methods .5 4.8.1 General. .5 4.8.2 Limit of quantification (LOQ) .5 4.8.3 Dynamic range .5 4.8.4 Determination of precision and trueness for quantitative methods .6 4.9 Robustness .6 4.9.1 General ..6 4.9.2 Robustness determination by interlaboratory study 4.9.3 Robustness determination bya multifactorial orthogonal test design ..6 5 Single-laboratory validation .6 6 Interlaboratory study (collaborative study) .7 6.1 General .7 6.2 Qualitative methods. 6.3 Quantitative methods .7 7 General laboratory and procedural requirements .7 7.1 General 7.2 Facilities, materials and equipment 7.3 Sample preparation and DNA extraction. .8 7.4 Use of controls. .9 7.5 Data analysis. 9 7.5.1 Control .9 7.5.2 Conventional PCR .10 7.5.3 Real-time PCR amplification curves .10 7.6 Expression of results. .10 7.6.1 Expression of positive results. .10 7.6.2 Expression of negative results ..11 7.6.3 Expression of quantitative results ..11 8 Test report. ..11 Annex A (informative) List of typical species used for inclusivity and exclusivity testing ..12 Annex B (informative) Examples of unit conversion methods from DNA copy numbers to theratioofmasses ..17 Bibliography .26 @ IS0 2019 - All rights reserved iii