人靶向核糖核酸酶(TR)酶联免疫elisa试剂盒
来自 : 蚂蚁淘
AimIncasesofgrosscontaminationthenakedeyemayidentifythepresenceofbacteriaandfungi.However,itisnecessarytodetectlow-levelinfectionsbyincubationofcellculturesand/ortheirproductsinmicroBIOLOGicalbroth.Equallythesesterilitytestscanbeusedtoconfirmtheabsenceofbacteriaandfungifromthepreparationwhichisimportantwhenpreparingcellbanksorcellcultureproducts.
Materials
- SoybeanCaseinDigest(TryptoneSoyaBroth,TSB)(15mlaliquots)(Prod.No.S1674)TSBPowder(Prod.No.T8907)
- FluidThioglycollateMedium(20mlaliquots)(TGM)(Prod.No.F4797)
- BacillussubtilisNCTC*
- CandidaalbicansNCTC*
- ClostridiumsporogenesNCTC*
Equipment
- Personalprotectiveequipment(latexmedicalgloves,laboratorycoat,safetyglasses)
- Waterbathsetto37ºC
- MicrobiologicalsafetycABInetatappropriatecontainmentlevel
- Centrifuge
- Incubatorsetat32ºC
- Incubatorsetat22ºC
ClickhereforFigure12.FlowSchemeforBacteriaandFungiTesting
Procedure
- Culturecelllineintheabsenceofantibioticsfor2passagespriortotesting.
- BringattachedcellsintosUSPensionwiththeuseofacellscraper.Suspensioncelllinesmaybetesteddirectly.
- Inoculate2xThioglycollateMedium(TGM)(Prod.No.F4797)and2xTryptoneSoyabroth(TSB)(Prod.No.T8907)with1.5mltestsample.
- Inoculate2(TGM)and2(TSB)with0.1mlC.albicans(containing100colonyformingunits,cfu).
- Inoculate2(TGM)and2(TSB)with0.1mlB.subtilis(containing100cfu).
- Inoculate1TGMwith0.1mlC.sporogenes(containing100cfu).
- Leave2(TGM)and2(TSB)un-inoculatedasnegativecontrols.
- Incubatebrothsasfollows:
- ForTSB,incubateonebrothofeachpairat32ºCtheotherat22ºCfor14days
- ForTGM,incubateonebrothofeachpairat32ºCtheotherat22ºCfor14days
- FortheTGMinoculatedwithC.sporogenesincubateat32ºCfor14days
- ExamineTestandControlbrothsforturbidityafter14days.
CriteriaforaValidResultAllpositivecontrolbrothsshowevidenceofbacteriaandfungiwithin14daysofincubationandthenegativecontrolbrothsshownoevidenceofbacteriaandfungi.
CriteriaforaPositiveResultTestbrothscontainingbacteriaorfungishowturbidity.
CriteriaforaNegativeResultTestbrothsshouldbeclearandshownoevidenceofturbidity.
Notes
- Thepositivecontrolsshouldbehandledinalaboratoryremotefromthemaintissueculturelaboratory.
- Controlorganisms(Bacillussubtilis,ClostridiumsporogenesandCandidaalbicans)arealsoavailablefromtheNationalCollectionofTypeCultures(NCTC),UK*.
- Thistestprocedureshouldbecarriedoutinamicrobiologylaboratoryawayfromthecellculturelaboratory.
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