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1、噬菌体的检查及效价测定1,purpose:1.1tounderstandthemeaningofphagetiteranditsdeterminationprinciple・1.2learnhowtocheckbacteriophages・1.3tomastertheoperationalskillsofdoublelayeragarplatemethodforphagetiterdetermination.2,principle:Bacteriophageisakindofviruswhichisspe
2、ciallyparasiticonbacteriaandactinomycetes,anditsindividualshapeisverysmal1.Itcannotbemeasuredbyconventionalmicrobialcountingmethod.Whenaphageinfectionbacteriawillcausecrackingsensitivebacteriarapidly,thereleaseoflargeamountsofprogenyphage,whichthenspreada
3、ndinfectionofthesurroundingcells,thesuspensioncontainingsensitivebacteriafromturbiditygraduallybecomesclear,orthenakedeyecanseetheplaque-plaqueinthetabletcontainingsensitivebacteria・Understendingthecharacteristicsofphages,rapiddetection,isolationandpotenc
4、ydeterminationareimportantforthepreventionofbacteriophagecontaminationinproductionandscientificresearch.Thesamplecanbefermentedliquid,air,sewage,soilandsoon.Asfortheobjectwhichcannotbesampled,thecottonsurfacesoakedwithsterilewatercanbeusedastheinspections
5、ample・Inordertoisolateeasily,thenumberofphagesinthesamplec^nbeincreasedbymultiplication.Phagedetectionusingbiologicalassayrequiresabout12h,sothatphagecontaminationcannotbedeterminedintime・Thephagecontaminationcanberoughlydeterminedbyrapidinspection,soasto
6、takenecessarycontrolmeasures・Accordingtothenormalfermentation(Culture)liquidaftercentrifugationwaslittleprecipitation,supernatantproteincontent,afterheatingisstillclearandinfectedwithphage;fermentation(Culture)activeproteinliquidaftercentrifugation,thesup
7、ernatantcontainingselfcleavingbacteriaescape,proteindenaturationoccursafterheating,andinthelightappearedandTyndalleffectnotclear・Thismethodissimple,rapidandaccurateinjudgingphagecontaminationinfermentationbroth・However,itisnotsuitableforthediagnosisofsolu
8、blebacteriaandphotosyntheticphage,anditisnotsuitablefortheprimaryseedculturemediumwhichislessinfectedbyphage・ThetiterofphageisthenumberofinfectivephagecontainedinImLsample・Doubleagarplatemethodwasusedtodeterminethet
