AOAC Official Method 999.09 VIP

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1、17.9.26Autoclaveat121°Cfor15min.Coolandadd2mL1%brilliantAOACOfficialMethod999.09greeendyesolution,(k),per1Lnonfatdrymilk.VIPforSalmonellafortheDetection(h)Brainheartinfusionbrothwithenrichmentsupplementcon-ofMotileandNon-MotileSalmonellainAllFoodstainingOxyraseÒ(BHI+O).Suspend37gcommercial

2、BHIin1LFirstAction1999deionizedwater.Mixthoroughly.Autoclavebrothat121°Cfor15min.Ondayofuse,add1mLenrichmentsupplementcontaining(ApplicabletoidentificationofSalmonellainallfoods.)OxyraseÒtoeach225mLbottleaftertestportionhasbeenaddedandmixingiscomplete.FinalpHis7.4±0.2.Caution:SeeAppendixB,

3、safetynotesonhandlingmicroorgan-(i)RappaportVassiliadisR10medium(RV).Suspend26.6gisms.DecontaminateallspentmediaandequipmentcommercialRappaportVassiliadisR10mediumin1Ldeionizedusedintestpriortodisposalofmediaorre-useofwaterandheatgentlytodissolve.Dispensein10mLaliquotsandequipment.steriliz

4、ebyautoclavingat116°Cfor15min.FinalpHis5.1±0.2.(j)5XRappaportVassiliadisR10medium(5XRV).SuspendSeeTable999.09Afortheresultsoftheinterlaboratorystudysup-133gcommercialRappaportVassiliadisR10mediumin1Lportingtheacceptanceofthemethod.deionizedwaterandheatgentlytodissolve.Dispensein5mLaliquots

5、in25´150mmtesttubesandsterilizebyautoclavingatA.Principle116°Cfor15min.Invisualimmunoprecipitate(VIP)assay,proprietaryantibodies,(k)Tetrathionatebroth(TT).Suspend46gcommercialTTwithahighspecificitytoSalmonellaantigens,areboundtobrothin1Ldeionizedwater.Mixthoroughly.Heatwithagitationchromog

6、eniccarrierand,separately,tosolidsupportmatrix.Re-andboilfor1mintocompletelydissolvepowder.Donotautoclave.agentsareincorporatedintestunitsandproducevisuallyCooltobelow45°C.Preparebrilliantgreendyesolutionbydissolv-discernablereactioninpresenceofSalmonellaspp.Duringinitialing1gdyeinsterilew

7、ateranddilutingto100mL.Add1mL1%hydrationoftestunit,Salmonellareactswithantibody-chromogenbrilliantgreendyesolutiontoTTbroth.Dispensein10mLaliquotscomplex.Antigenantibody-chromogencomplexisformed,whichinsteriletesttubesandstoreat48°C.PrepareIKIsolutionbydis-flo