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MLT-BiochemistryTest

Microbiology

QuestionAnswer
Coagulase test is used to? To differentiate Staphylococcus aureus from other staphylococcus species.
How does coagulase work? Coagulase turns fibrinogen to fibrin(a visible clot).
What is staphylokinase? It can breakdown fibrin. So the coagulase test has to be examined after 4 hrs, but not more than 24 hrs.
How does Urease test work? It turns Urea to Ammonia, with indicator Phenol red turns to bright red.
Condition for Urease test? 18-24hrs at 37 degree celsius. PTC: Proteus spp. NTC: E.coli
what is Beta-galactosidase (ONPG) test? ONPG structure is chemically similar to galactose. It will be broken down by beta-galactosidase to Orthonitrophenol (yellow in colour) and galactose. For: Shigella sonnei PTC: E.coli NTC: Proteus spp.
Nitrate reductase test? To test the presence of nitrate reductase, Nitrate to Nitrite. For: some Enterobacteriaceae. Incubate the organism with nitrate broth overnight. Add 1ml of reagent A and 1ml of reagent B. Positive: Red. Negative: yellow. Confirmed with Zinc( to Red).
PTC and NTC for Nitrate reductase test PTC: E.coli NTC: Actinobacter baumanii
Oxidative /Fermentation of glucose, O/F test is used for ? To determine the way of sugar break down , which could be oxidation, fermentation or unable to utilise sugar.
What is O/F medium? Content? Hugh and Leifson medium. Low protein: CHO ratio semi-solid consistency to allow acid formed to permeate to the medium from surface. Bromothymol blue as pH indicator, turn yellow when acidic.
How does O/F test work? Prepare each test organism two tubes with O/F medium, one with paraffin oil on top. stabbing deep into the agar medium with a straight wire. Yellow-Green: Oxidative(Pseudomonas); Yellow-Yellow: Fermentative(E.coli);Green-Green: Asacharolytic(Alcaligenes)
What is Aseculin Hydrolysis? Aesculin will be broken down to Aesculetin and glucose. Asesculetin with Ferric citrate will form dark colouration.
Aseculin test is used for? Only Group D streptococci(Enterococci) can hydrolysis aesculin in the presence of bile salt. PTC: Enterococcus faecalis NTC: streptococcus pyogenes
What is Kovac's reagent? It indicates the formation of Indole.
How does Indole form? Tryptophan will be broken down by tryptophanase to form indole, ammonia, and pyruvate.
PTC and NTC for indole-Kovac's test PTC: E.coli NTC: Klebsiella
Optochin sensitivity test? To identify streptococcus pneumoniae from other alpha-hemolysis streptococci spp. Blood agar, incubate at 5% CO2, then add Optochin disc. Sensitive: Streptococci pneumoniae; resistant: Viridans streptococci
What is alpha- and beta-hemolysis? alpha hemolysis: partly hemolysis of RBC beta hemolysis: completely hemolysis of RBC
Bacitricin sensitive test? To identify streptococcus pyogenes from other beta-hemolysis streptococci. Sensitive: Streptococcus pyogenes(Group A) Resistant: Streptococcus agalactiae( Group B)
Created by: kencho