第1页：Isolation and identification
Isolation and identification
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第2页：Objectiveof Experiment
Objectiveof Experiment
- To masterprimaryprincipleandmethodofisolation and identificationpyogeniccoccifrom clinicalspecimens.
- Todiagnose clinicaldisease and guide usto use medicines .
第3页：Specimens(Gram-stain)Morphological
Specimens(Gram-stain)Morphological
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第4页：Identified method for
Identified method for
- Gram-stain
- Isolationand culture
- Pure culture
- Directidentification
- The antibiotic susceptibility test
第5页：Gram-stain
Gram-stain
- Morphological characteristics-Typical staphylococcus cells arespherical,arranging in irregularclusters,which just like as clusters of grape.–Typical cells are Gram-positive,nonmotile and do not formspore.–The cell is about 1μm in diameter.
第6页：Staphylococci are
Staphylococci are
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第7页：Isolationand culture
Isolationand culture
- Cultural–Specimens planted on blood agar platesgive rise to typical colonies in 18 hours at370C–Hemolysis and pigment production may notoccur until several days later and areoptimal at room temperature.
第8页：Isolationand culture
Isolationand culture
- colonial characteristics–Colonies of staphylococci on solid meida areround,smooth,raised,and glistening.S.aureusforms gray to deep golden yellow colonies.–S.epidermidis colonies are gray to white onprimary isolation.–Various degrees of hemolysis are producedby S.auerus and occasionally by otherspecies.
第9页：
第10页：Pure culture
Pure culture
- MATERIALS:–Agar slope culture–Colonies on agar plate
- PROCEDURE–Withtheflame-sterilizedwireinoculatingloop,transferasmallamountofbacteriafromthecolonyonagarplate.Thenstreakontheagarslope.–Sterilethemouthoftubes,replugthetesttubesandflametheloop.–Labelandincubateat37℃for18-24hours
- .
第11页：Directedidentification
Directedidentification
- The Coagulase Test
- The Dnase Test
- The mannitol fermentation test.
- The phage typing test
- Animal Experiment
第12页：The Coagulase Test
The Coagulase Test
- Possession of the enzyme coagulasewhich coagulase plasma is an almostexclusive property of Staph.aureus.Thereare two ways of performing this test–The Slide Coagulase Test–The Tube coagulase Test
第13页：The Coagulase Test
The Coagulase Test
- Coagulase is an enzyme convertingfibrinogen into fibrin promoting bloodclotting. It has been speculated thatthis enzyme might be a virulence factorwith the coagulated blood around thebacteria protecting them from the immunesystem.
- However, coagulase-negative strains areoften as pathogenic as coagulase-positive strains.
第14页：The Slide Coagulase Test
The Slide Coagulase Test
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第15页：The Tube coagulase Test
The Tube coagulase Test
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第16页：The Dnase Test
The Dnase Test
- Inoculate Dnase agar plates with a loop so thatthe growth is in plaques about 1 cm indiameter.Incubate at 370C overnight.Flood theplate with 1 N hydrochloric acid.Clearingaround the colonies indicates Dnaseactivity.The hydrochloric acid reacts withunchanged deoxyribonucleic acid to give acloudy precipitate.A few other bacteria,e.g.Serratia,may give a positive reaction.
第17页：The mannitol fermentation
The mannitol fermentation
- Inoculate the bacteria into a mannitolmicro-tube,incubate at 370C for18h.S.aureus will ferment mannitol toproduce acid,which causes the medium toturn yellow.
第18页：The phage typing test
The phage typing test
- This test is used to trace the infectiveagent in epidemiology if necessary.It isusually not done for routine clinicalpurpose.
第19页：Animal Experiment
Animal Experiment
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第20页：The antibiotic
The antibiotic
- This test is helpful for the treatment ofS.aureus infection.
- materials–S.aureus（isolated from the pus of apatient）.–Several kinds of filter paper（each containsdifferent kinds of antibiotics）–Nutrient agar plate
第21页：The antibiotic susceptibility
The antibiotic susceptibility
- PROCEDURE–StreakingtheS.aureusonagarplate(thoroughlycoveredtheplate)–Put 4 kinds of paper contained differentantibiotics on the plate (each paper are far awayabout 2 cm)–Incubate ar 370C 18-24 hours.–Observetheresultstheplatesareexaminedforthepresentofzonesofinhibitionofbacterialgrowtharoundthefilterpaper.Thesensitivityoftheorganismisindicatedbythediameterofthezoneofgrowthinhibition.
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第23页：Dimidiation the
Dimidiation the
- Lactasefermenters:saprophyticandcommensalEscherichiaKlebsiellaCitrobaterEnterobacterEnterobacter,
- Nonlactasefermenters:pathogensSalmonellaShigellasomeCitrobacter,ProteusSerratia
第24页：Biochemical reactions of
Biochemical reactions of
- SpeciesbuttslopeH2SmotilityE.coliAGAG-+SalmonellaA-+/-+OtherSalmonellaAG-+/-+ShigellaA-+/--butt:fermentdextroseslope:lactose
- A:acidAG:acidandgas
第25页：Colonialcharacteristicobservation
Colonialcharacteristicobservation
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第26页：Specimens
Specimens
- Different specimens should be taken dependingon the kind and the process of the disease.bloodbone marrowUrinestool
第27页：Isolation
Isolation
- Culture medium: S.S agar
- Method: streak plate
- Result:unpathogenetic colonies: middle size, redSuspect colonies: colorless, small, opaque
第28页：Escherichia coli
Escherichia coli
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第29页：Shigella sonnei
Shigella sonnei
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第30页：Salmonella
Salmonella
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第31页：Vibrio cholerae
Vibrio cholerae
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