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1、17.10.04C.ReagentsAOACOfficialMethod993.09(Caution:ProbesolutionandpositivecontrolsolutioncontainListeriainDairyProducts,Seafoods,andMeats0.1%sodiumazide.DisposalofthisreagentintosinkswithcopperColorimetricDeoxyribonucleicAcidHybridizationMethodorleadplumbingshouldbe
2、followedimmediatelywithlargequanti-(GENE-TRAKListeriaAssay)tiesofwatertopreventpotentialexplosionhazards.)FirstAction1993Storepretreatmentreagentconcentrate,lysisreagentconcentrate,FinalAction1996probesolution,enzymeconjugate100´concentrate,sub-stratechromogensolutio
3、n,positivecontrolsolution,andnegative[MethodistestprocedureforpresenceofListeriaspeciesindairycontrolsolutionat28°C.Storeallothersolutionsanddipsticksatproducts,meats,andseafoods.Becausecertainpercentageoffalse-roomtemperature(<30°C).positivereactionsmaybeencountered
4、,positiveassaysmustbecon-(a)Pretreatmentreagentconcentrate.150mgLysozymeandfirmedbystandardculturemethods(seeH).]3000unitsmutanolysinin0.1Mpotassiumphosphatebuffer.(b)Pretreatmentreagentbuffer.0.1MTrispH7.17.4,SeeTable993.09fortheresultsoftheinterlaboratorystudysup-0
5、.01Mdisodiumethylenediaminetetraacetate(EDTA),andportingtheacceptanceofthemethod.0.0075%bromphenolblue.A.Principle(c)Lysisreagentconcentrate.SerineproteasederivedfromDetectionofListeriaribosomalRNA(rRNA)usesspecificDNATritirachiumalbum(ProteinaseKissuitable).probes.F
6、ollowingprimaryandsecondaryenrichmentoftestpor-(d)Lysisreagentbuffer.5%n-lauroylsarcosine(Sarkosylistions,bacteriaarelysed,andlabeledListeria-specificDNAprobessuitable),0.005MEDTA,0.26MTrispH7.27.6,1.0MNaCl,andareaddedforsolutionphasehybridization.IfListeriarRNAispre
7、s-0.05%brilliantyellow.entintestportion,fluorescein-labeleddetectorprobeand(e)Listeriaprobesolution.Fluorescein-labeled,Listeria-spe-polydeoxyadenylicacid(polydA)-tailedcaptureprobewillhybrid-cific,syntheticoligonucleotideDNAprobeandpolydeoxyadenylicizetotargetrRNAse
8、quences.Apolydeoxythymidylicacid(polyacid(dA)-tailed,Listeria-specific,syntheticoligonucleotideDNAdT)-coatedplasticdipstick(solidph
