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1、17.10.07prepared8MLiClstocksolution(Sigma),add2.65mLper225mLAOACOfficialMethod996.14sterilizedmFB.DetectionofListeriamonocytogenes(i)BufferedListeriaenrichmentbroth(BLEB).—Suspend36.1gandRelatedListeriaSpeciesinSelectedFoodscommercialListeriaenrichmentbrothin1
2、Lwater.Add8.5gandfromEnvironmentalSurfaces3-(N-morpholino)propanesulfonicacid(MOPS)and13.7gAssurancePolyclonalEnzymeImmunoassay(EIA)MOPS–Nasalt.Mixwellandheattodissolveifnecessary.DispenseFirstAction1996in9mLaliquots.Sterilizebyautoclavingat121°Cfor15min.Revis
3、edFirstAction1999Items(a)–(g)areavailableasAssuranceListeriaEnzymeFinalAction2001Immunoassay(EIA)testkitfromBioControlSystems,Inc.(12822Revised2001SE32ndSt,BellevueWA98005USA).(ApplicabletodetectionofListeriamonocytogenesandrelatedListeriaspeciesindairyfoods,r
4、edmeats,pork,poultryandpoultryC.Apparatusproducts,seafood,fruits,vegetables,nutmeats,pasta,chocolate,(a)Incubators.—Maintaining30–32°Cand35–37°C.eggs,bonemeal,andfromenvironmentalsurfaces.)(b)Waterbath.—Maintaining100°C.Alternatively,flowingCaution:L.monocytog
5、enesinfectionscancausefetaldeath.Itissteamautoclavesetat100°Coradryheatblockmaybeused.recommendedthatpregnantwomenavoidhandlingthis(c)Microplatewasherorplasticsqueezebottle.—Forwashingorganism.Sterilizecontaminatedequipmentandmediamicrowellstrips.beforedisposa
6、lorreuse.(d)Microplatereader.—Photometerwith405–410nmfilter,readingmicrowellplates.Mayincludeoptionalprinter.SeeTable996.14fortheresultsoftheinterlaboratorystudysup-(e)Micropipets.—Accuratelydispensing0.1and1.0mL.portingacceptanceofthemethod.(f)Vortexmixer.—Fo
7、rmixingtestportions.(g)Toploadingbalance.—Forweighingtestportions,measur-A.Principleingupto1000g,sensitivyof±0.1g.Proprietaryantibodieswithahighspecificityto(h)Blenderorstomacher.—Forhomogenizingtestportions.L.monocytogenesandrelatedListeriaspeciesantigensareb
8、oundtomicrowellplates.EnrichedtestportionsandpositivecontrolsareD.GeneralInstructionsaddedtotheplate.Anyantigensthatarepresentbindtothemicrowells,formingantibody-antigencomplexes.N