ISO INTERNATIONAL STANDARD 21702 First edition 2019-05 Measurement of antiviral activity on plastics and other non-porous surfaces Mesure de I'activite antivirale sur les matieres plastigues et autres surfaces non poreuses Reference number ISO 21702:2019(E) ISO @ IS0 2019 IS0 21702:2019(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 21702:2019(E) Contents Page Foreword ..V Introduction. ...vi 1 Scope. 2 Normative references 3 Terms and definitions 4 Materials .2 4.1 Virus and host cell to be used for the tests .2 4.2 Reagents .2 4.3 Culture medium and solutions. 2 .2 4.3.1 General. 4.3.2 Eagle's minimum essential medium (EMEM) 2 4.3.3 RPMI 1640 medium .2 4.3.4 7,5 % sodium bicarbonate solution .3 4.3.5 Formaldehyde solution. .3 4.3.6 Methylene blue solution .3 4.3.7 Inactivated fetal bovine serum (FBS) 3 4.3.8 Growthmedium. .3 4.3.9 Maintenance medium 3. 4.3.10 Double concentration of the maintenance medium ..4 4.3.11 Phosphate buffered saline [PBS (-)] 4 4.3.12 Trypsin derived from beef pancreas and PBS (-) solution .4 4.3.13 Trypsin EDTA solution. 4 4.3.14 DEAE-dextran solution ..4 4.3.15 Agar medium for plaque assay .5 4.3.16 Soybean casein digest broth with lecithin and polyoxyethylene sorbitan monooleate (SCDLP broth) ..5 5 Apparatus ..5 6 Preparation ..6 6.1 Restoration of host cell from cryopreservation ..6 6.2 Subculture of host cell 1 6.3 Cell culture for measuring virus infectivity titer .7 6.4 Preparation of test inoculums. .7 6.4.1 Influenza virus. .7 6.4.2 Feline calicivirus. 6.5 Preparation of test specimens ..8 6.6 Controltest. ..9 6.6.1 General .9 6.6.2 Verification of cytotoxic effect on host cell 9 6.6.3 Verification of cell sensitivity to virus and the inactivation of antiviral activity.... 9 7 Test procedure .11 7.1 Preparation of test specimen. .11 7.2 Inoculation of test specimens .11 7.3 Contact of the inoculated test specimens. .12 7.4 Recovery of virus from test specimens. .12 7.4.1 Test specimens immediately after inoculation ..12 7.4.2 Test specimens after contact. ..12 7.5 Determining the infectivity titer of virus by plaque assay ..12 Expression of results. ..13 8 8.1 Determination of the infectivity titer of virus ..13 8.2 Conditionsforavalidtest .13 8.3 Calculation of the antiviral activity .14 @ IS0 2019 - All rights reserved iii