第1页：Isolation and identification
Isolation and identification
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第2页：Objectiveof Experiment
Objectiveof Experiment
- To masterbasicprincipleandmethod,which use to isolate and identifypyogenic coccifrom clinicalspecimens.
- Todiagnoseclinicaldisease and guidedoctors to select proper 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
第5页：Staphylococci are
Staphylococci are
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第6页：
第7页：Directidentification
Directidentification
- The mannitol fermentation test.
- The Coagulase Test
- The Dnase Test
- The phage typing test
- Animal Experiment
第8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 themedium to turn yellow.
第9页：Test of Microbiological
Test of Microbiological
- While a patient is infected by pathogenicenterobacteria (Salmonella),how to diagnose withmicrobiological methods in lab?(simple procedure)
- While a patient is infected by pyogenic cocci,howto diagnose with microbiological methods inlab?(simple procedure)
第10页：The Coagulase Test
The Coagulase Test
- Coagulase is an enzyme converting fibrinogeninto fibrin promoting blood clotting.
- It might be a virulence factor with thecoagulated blood around the bacteria protectingthem from the immune system.
- Coagulase-negative strains are often aspathogenic as coagulase-
第11页：The Slide Coagulase Test
The Slide Coagulase Test
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第12页：The Tube coagulase Test
The Tube coagulase Test
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第13页：The Dnase Test
The Dnase Test
- Inoculate Dnase agar plates with a loop sothat the growth is in plaques about 1 cm indiameter.Incubate at 370C overnight.Floodthe plate with 1 N hydrochloricacid.Clearing around the coloniesindicates Dnase activity.The hydrochloricacid reacts with unchangeddeoxyribonucleic acid to give a cloudyprecipitate.A few other bacteria,e.g.Serratia,may give a positive reaction.
第14页：The phage typing test
The phage typing test
- This test is used to trace the infectiveagent in epidemiology if necessary.
- It is usually not done for routineclinical purpose.
第15页：Animal
Animal
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第16页：The antibiotic susceptibility
The antibiotic susceptibility
- This test is helpful for the treatmentof S.aureus infection.
- materials
- Staphaureus（isolated from the pus ofa patient）.
- Several kinds of filter paper（eachcontains different kinds of antibiotics）
- Nutrient agar plate
第17页：The antibiotic susceptibility
The antibiotic susceptibility
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第18页：
第19页：Laboratory Diagnosis of
Laboratory Diagnosis of
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第20页：Dimidiation of the
Dimidiation of the
- Lactasefermenters:saprophyticandcommensalEscherichiaKlebsiellaCitrobaterEnterobacter
- Nonlactasefermenters:pathogensSalmonellaShigellasomeCitrobacterProteusSerratia
第21页：
第22页：Colonialcharacteristicobservation
Colonialcharacteristicobservation
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第23页：Specimens
Specimens
- Different specimens should be takendepending on the kind and the process ofthe disease.bloodbone marrowUrinestool
第24页：Isolation
Isolation
- Culture medium: S.S agar
- Method: streak plate
- Result:Pathogenetic colonies: middle size, redSuspect colonies: colorless, small,opaque
第25页：Biochemical reactions of
Biochemical reactions of
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第26页：Antigens of salmonella
Antigens of salmonella
- O antigenpolysaccharideof LPSsomatic antigenused to divide Salmonella to 42 groupsA-Z groups are pathogenicstable to heat (remains activity at 100℃)H antigenflagella antigendivide Salmonella tow phase:special no specialVi antigenrelated to the virulence of Salmonellasensitive to heat ( lose activity at 60℃)
第27页：
第28页：Salmonella
Salmonella
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第29页：Serological Identification of
Serological Identification of
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第30页：Gram Stain
Gram Stain
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第31页：Gram stain of Nocardia
Gram stain of Nocardia
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第32页：flagella
flagella
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