‘cote cman gee 7 PVC ‘Altornative methods for agribusiness ‘Analytical porformances certified VALIDATION CERTIFICATE FOR ALTERNATIVE ANALYTICAL METHOD [ACCORDING TO STANDARD EN ISO 16140: 2003 ‘Gotieate No CHR FINA 1206 ‘Validation dat: “a2006 Renewal dates: 28.00.2009, ‘s02013 End of vay: sai22017 ‘The Company — CHRONagar ‘Ste of cHromagar (Head otic)” 4, placed 18 jun 1340 production 60-52 avenue de See ‘6008 Pare Franco ‘75018 Pars France Reta Laboratoires HUMEAU. ‘rue Kaplor Ade Gesvine ~ 9° 4125 4241 La Chapole-su-Ertre Codex —France is hereby authorized to rofer to the NF VALIDATION certifate for the folowing atematve ‘quanttave analysis meiod CHROMagar™ Listeria numeration Pretocolreference: CHROMags Listeria: NT-EXT.009- Version 5 (Grrtonagar™ lertcaton Lister: NT-EXT-025 ~ Version 4 sore ‘A human font an production entonmant samples. ResTRICTIONS None REFERENCE METHOD EN 180 11280-2 (August 1999) and ts amendment At (February 2008) Food tlccbiabay — horizontal mahod forthe delacon and enumeraton e Listeria manoeyogenas «Patt 2: Enumeration i £ Managing Director Florence MEAUX HER ‘AFWOR Cariienton “ve Fai de Presson 0671 Laan Slit Dels Coon Fanon one sh G99 00 ro a0 17000, i218 PRINCIPLE OF THE METHOD CCHROMagar™ Listeria numeration for enumeration of Lsiors manceylaganes fs a matted cutng 2 fuepersion ban andthe ekemogenleesture media GHROMager™ ora, alow to spose ‘elecon of Lstona monoeloganes. In caoo of postive result wit tho enumeration method, conan is ot necessary when the Presence of Listeria moneeyogenes has already beon covfmod bythe deacon method In te ater cases, samples ienited as postive by the ateratve method must be contre by one ofthe folowing means! ‘+ Accorting to casical toss descr in methads standardised by CEN or ISO (nclidna Dareaton Sp) staring torn CHROMaga™ Litera ‘+ Diecty ram a ypica colony wel olted on CHROMagar™ Listeria by spoing the colony onto CCHROMagar™ Idenieaon storia’ Lista. manceylogenes “deplays =) mauve” clout Surounded by a white opaque hab. Noto contimaton has aeady been done fora same sample ding the detection phase win the CChkoMagar Lista mathos (CHR 2101" 12101), anew contemavon not necessary atthe ‘CHoMagar Lists enumeraton method you do not conf 8 cxiones counting, can crate a isk of making a overestimated resut becouse of the possible prosance of pial colonies that weuld note. manasyfogenes In te event of discordant resus (presumptive postive with aerate method, non-comfnss by ‘means f options described above) the labora mus! flow te necessary slaps to ene vay of theresutsobainea NOTE (validation history) In Sepemiber 2009, the carte was renewed for CHROMagar™ Listeria numeraton vithout ‘complementary assays. Sie the prevcus valaton In 2005, the formula of CHROMagar™ Utes rumoraton tat haa A changed, ob well reerenee moto and te EN ISO T6140 prfoco, In October 2013, the carteation cf he aternave method hasbeen renewed This ones not change ‘shoo te lst valdaton sty, ana also te reference maths. The hertaboratory resute cbtaiies 2008 wore re analyeedaezading o EN ISO TBIAUIAT, wanout any npst ont study conclusion Tests ware dene with two plats by luton forthe erence mathe, and ane plate by don fhe ‘Sernave msios LINEARITY AND relative ACCURACY ‘Comparison of performances ofthe altrative method and the reference method Linearity study: Tests were pavormed i 2008 un Ue 5 “oud producti conesains se fr the fd atogotes gen he tabla below ‘The samples wore anayzed in duplieate vith each ofthe two methods, atthe fe faving atin cotaminaten levels: 101050, 80 10 100, 100 to 600, 600 to 1 000 and 1/000 to 13000 crus ‘The folowing results were obtane:38 Food category Food produtsrain pair | Regresion ine | ‘iat prsves Pate 7L monogjoperes de Y= 0582 X+ 0598 Day rods Faw nik moncofogenes ia __|Y=1011X=004 [vegetal products |Salad monocytogenes 2a ‘Ye 024x005 Seafood products | Smoked salmon, monoaytoganes tiie | ~Y="1006 x= o0x1 Envteoment samples [instil watr/L. monocytogenes ti2a | Y= 0.967 X + 0.122 YY log sttative mato) OG roterence meta) Accuracy study Tests wore performed in 2008. The stata! interpretation was condita on 84 rasuts, acusing 10 natraly contaminated samples and 44 rtfttly contaminated samples, belonging © the folowing major ood catogores: oat products, daly products, vegetal products, seafood grits and envionment samples ‘The samples wee analyzed in duplicate wth each of the two methods, ‘Asan nication the cortamnation (concanation) ranges wore flows Food category [Contamination range (in fog CFU eat products - 17010415, Day products 114810406, - ‘Vege roduc 1,500 400, Seafood products "1g4105 11 Enveonment saps 47010818 id ‘Tho equaton ofthe repression Ine betwean the aternatve method and he reference mated, for aleatagores combined 8 flows Y= 1908 X- 0012 Y stag sttmatve matoa) log roteronce metred) ‘The repeataity standard devaton fr both msds andthe bas between the two methods were Aoterminad acceding te the msod of caleton uted forthe nierisborsony sty (ea sans 1.35 and 6.36 of he standard EN ISO TeT40/AT). Those results prove addtional normal or he accuracy extern “Te repeataity standard devaton (ino) obtained forthe aenative method and he eleence mathe ae a lows ‘Atmative method Reforanca metios ‘Sra O18 ‘Sret'= 0900 NB Limit repatatity ¢=2.8 5, wh; repeat standard devon “Th bias (ng) tetwoun the to metiods(aemative method - eerence method) a alors D=0,02 one consider the mean valve ‘P= 0,01 if ene considers the averap of inva bias. CConetusion for neat and relative accuracy Stiles of wary and reatine accuracy show that results abained withthe strnabve metic are Simla those obtained wih the reference mothod ‘SSO OBC CFM CR EH is oS