革蘭氏陽性球菌的分離與鑒定第1頁，課件共27頁，創作于2023年2月IdentifiedmethodforStaphylococciGram-stain

Isolationandculture

Pureculture

Direct

identification第2頁，課件共27頁，創作于2023年2月StaphylococciareGram-positivecocci,typicallyarrangedinclumpsorGrape-likeclusters第3頁，課件共27頁，創作于2023年2月第4頁，課件共27頁，創作于2023年2月Direct

identification

Themannitolfermentationtest.

TheCoagulaseTest

TheDNaseTestNovobiocin(NB)disc第5頁，課件共27頁，創作于2023年2月Themannitolfermentationtest.

Inoculatethebacteriaintoamannitolmicro-tube,incubateat370Cfor18h.S.aureuswillfermentmannitoltoproduceacid,whichcausesthemediumtoturnyellow.第6頁，課件共27頁，創作于2023年2月TestStaphylococcusaureusStaphylococcusepidermidisStaphylococcussaprophyticusMannitolfermentationPositiveNegativeUsuallypositive第7頁，課件共27頁，創作于2023年2月TheCoagulaseTestCoagulaseisanenzymeconvertingfibrinogenintofibrinpromotingbloodclotting.Itmightbeavirulencefactorwiththecoagulatedbloodaroundthebacteriaprotectingthemfromtheimmunesystem.Coagulase-negativestrainsareoftenaspathogenicascoagulase-positivestrains.第8頁，課件共27頁，創作于2023年2月TheDNaseTestInoculateDNaseagarplateswithaloopsothatthegrowthisinplaquesabout1cmindiameter.Incubateat370Covernight.Floodtheplatewith1Nhydrochloricacid.ClearingaroundthecoloniesindicatesDNaseactivity.Thehydrochloricacidreactswithunchangeddeoxyribonucleicacidtogiveacloudyprecipitate.Afewotherbacteria,e.g.Serratia,maygiveapositivereaction.第9頁，課件共27頁，創作于2023年2月StaphylococcusaureusGrowingonDNaseAgar

第10頁，課件共27頁，創作于2023年2月Bloodagarwithanovobiocin(NB)disc

StaphylococcusaureusGrowingonBloodAgar

StaphylococcussaprophyticusGrowingonBloodAgar第11頁，課件共27頁，創作于2023年2月IdentificationofGram-positivecocci第12頁，課件共27頁，創作于2023年2月typesOfStreptococcus第13頁，課件共27頁，創作于2023年2月βhaemolyticsteps:LancefieldgroupsHistoricallyA,B,C,FandGgroupspecificantigenscellwallpolysaccharidegroupDandEnterococcilipoteichoicacidmoietyNotalwaysβhaemolyticNow:18differentgroupsofstrep:fromA-H,andfromK-T第14頁，課件共27頁，創作于2023年2月(β)HaemolyticStreptococcusgroupsS.pyogenesGroupA

S.agalactiaeGroupB(occasionallyαornone)

StreptococcusequismilisGroupC(occasionallyαornone)

StreptococcusbovisGroupD(αornone,occasionallyβ)

StreptococcusanginosusGroupF&G第15頁，課件共27頁，創作于2023年2月GpAstrep:S.pyogenesLaboratoryIDGram+veCatalaseHaemolysissmall,round,greycoloniesβhaemolysiszoneislargeGpAantigen第16頁，課件共27頁，創作于2023年2月IsolationandIdentificationofgroupAbetahemolytic(Streptococcuspyogenes)

onBloodagar.Streptococcuspyogenesproducesazoneofbetahemolysisaround2-3mmindiametersurroundingeachcolonySensitivitytotheantibioticbacitracinGroupAStrepissenstive,otherbhaemolyticstrepsareresistant.第17頁，課件共27頁，創作于2023年2月Bacitracinsensitivity第18頁，課件共27頁，創作于2023年2月GroupB:S.agalactiaeS.agalactiaeIdentificationCatalasenegativeAppearanceonBALargercoloniesthanGpASmallzoneofhemolysisLatexagglutinationGpBBacitracinresistantCAMPpositiveGrowthonMac(weak)第19頁，課件共27頁，創作于2023年2月PresumptiveIDTests-CAMPThe“CAMP”test(acronymfordevelopers)isusedtodifferentiateS.agalactiae(GBS),whichispositive,fromotherbetahemolyticstrep,allofwhicharenegative“CAMPfactor”isasolublehemolysinproducedbyGBSthatcombinesinasynergisticwaywithasimilarhemolysinofS.aureustoformanarrowheadzoneofclearing.SeeimageThestaphisstreakedperpendicularlytotheunknownstrep.Afterappropriateincubationtimean“arrowhead-shaped”clearzoneofhemolysiswillappear.第20頁，課件共27頁，創作于2023年2月S.agalactiaeSynergistichaemolysisobservedbetweenbetahemolyticStaphylococcusaureusandgroupBstreptococci.Positivereaction:arrowheadhaemolysispattern第21頁，課件共27頁，創作于2023年2月α–Haemolyticstreps:ViridansstrepsNotgroupablebyLancefieldgroupsNogroupspecificCHO

StreptococcuspneumoniaePrimaryhumanpathogenS.sanguis

S.mitis

S.mutans

S.salivarius第22頁，課件共27頁，創作于2023年2月第23頁，課件共27頁，創作于2023年2月ThePneumococcus(Streptococcuspneumoniae)

1:IsolationonBloodagar

PneumococcifrequentlyrequireenrichedmediaandincreasedCO2tensionforinitialisolation.TheyareusuallyisolatedonBloodagarandincubatedinacandlejar(aclosedcontainerinwhichalitcandleisplacedtoremoveO2andincreaseCO2)at37C.OnBloodagar,coloniesappearsmall,shiny,andtranslucent.Theyaresurroundedbyazoneofalphahemolysis

2:Optochinsensitivity

Pneumococciaretheonlystreptococcithataresensitivetothedrugoptochin(ethylhydrocupreinehydrochloride).3:Gramstain:

gram-positive,diplococci

第24頁，課件共27頁，創作于2023年2月第25頁，課件共27頁，創作于2023年2月TheGenusEnterococcus

Enterococciaregram-positivestreptococci,typicallyoccurringinpairsandshortchains,thatarenormalfloraoftheintestinaltract.Enterococciresponsibleforavarietyofopportunisticinfectionsinhumans,andserologicallybelongtoLancefieldgroupDstreptococci.OnBile