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1、17.10.05Monoclonalantibodiesintestkitmustreactwithanepitopecom-AOACOfficialMethod994.03montoallspeciesofListeria,butnotpresentinotherGram-positiveListeriamonocytogenesinDairyProducts,organisms.Seafoods,andMeats(b)Controlantigens.(1)Positivecontrol.Lyophilized,puri-Colorimet
2、ricMonoclonalEnzyme-LinkedImmunosorbentfiedListeriaantigenreactivetoListeriaspp.antibodies.(2)Nega-AssayMethod(Listeria-Tek)tivecontrol.Lyophilized1%nonfatdrymilk.BothcontrolsFirstAction1994preservedwith0.1mg/mLgentamicinsulfate.StorereconstitutedFinalAction1997antigensat28
3、°Cfor60days.(c)Conjugateandconjugatediluent.Anti-Listeriaantibodies(ProcedureforpresenceofL.monocytogenesindairyproducts,(murine)conjugatedtocommerciallyavailablehorseradishseafoods,andmeats.Testisnotconfirmatorybecausemonoclonalperoxidaseinliquiddiluentwithproteinstabilize
4、rsandantibodiesusedintestmaycross-reactwithothermembersofthege-antimicrobialagentinTrisbufferpH7.6.MonoclonalantibodiesinnusListeria.)conjugatemustreactwithepitopecommontoallListeriaspecies,Enrichmentbrothsfromtestportionspositivebythistestmustbebutnotreactwithepitopeadsorb
5、edtomicrotiterstripplates.streakedonselectivemediaasin992.18C(see17.10.02)and(d)TMBperoxidasesubstrate(SolutionA).3,3¢,5,5¢-992.19CandE(see17.10.03).TypicalorsuspiciouscoloniesmustTetramethylbenzidine,0.4g/L,inaproprietaryorganicbase.beidentifiedbiochemicallyasL.monocytogen
6、es.(e)TMBperoxidasesubstrate(SolutionB).CitricacidbufferPositiveresultisvalidonlywhennegativeandpositivecontrolscontaining0.02%H2O2.showacceptableabsorbancereadings.(f)Bufferedwashconcentrate(25´).AqueousmixtureofSeeTable994.03forinterlaboratorystudyresultssupportingthe19.2
7、5%NaCl,1.25%Tween20,0.525%KH2PO4,andacceptanceofthemethod.1.825%K2HPO4.A.Principle(g)Stopsolution.1Msulfuricacid(H2SO4).Listeriaantigensaredetectedbasedonenzymeimmunoassay(h)ModifiedFraserbroth.5.0gpancreaticdigestofcasein,(EIA)usingspecificmonoclonalantibodies.Polystyrenem
8、icrotiter5.0gpepticdigestofanimaltissue,5.0gbeefextract,5.0gyeastex-wellsarecoated
