Hydrogen peroxide is often used as a topical disinfectant in wounds, and the bubbling that is seen is due to the evolution of o 2 gas. The slide coagulase test may yield a negative result for. Coagulase test principle, procedure and result interpretation. Place a drop of coagulase plasma preferably rabbit plasma with ethylenediaminetetraacetic acid edta on. Do not use this procedure for mycobacteria testing. Taylor and mcdiarmid 18 used rabbit, sheep, and ox plasmas in the plate test. I watched many variants of the procedure on youtube. Place a drop of distilled water or saline next to the drop of plasma as a control. Be sure that you have a staph before running this test. The coagulase test for staphylococci sciencedirect. Why should the test tube coagulase test be incubated for no more than 24 hours. Coagulase production is confined to pathogenic strains of staphylococci and this test is specific for the identification of these strains.
Coagulase test is used to differentiate staphylococcus aureus positive from coagulase negative staphylococcus cons. Coagulase test principle, procedure and result interpretation purpose of coagulase test. Catalase serves to neutralize the bactericidal effects of hydrogen peroxide. Therefpre, tube coagulase test still remain a test of choice for s. How soon will agglutination appear on the coagulase slide test if it is a positive result. With the loop, straight wire or wodden stick, emulsify a portion of the isolated colony in each drops to make two thick suspensions. Intended use the bbl staphyloslide latex test is a latex slide agglutination test for the differentiation of staphylococci which possess clumping factor andor protein a, usually. Bound coagulase is detected by the slide coagulase test, whereas free coagulase is detected by the tube coagulase test. Catalase test principle, uses, procedure, result interpretation with precautions. The enzyme coagulase is demonstrated invitro by two methods a the slide coagulase test b the tube coagulase test.
Free coagulase reacts with a substance in plasma to form a fibrin clot. Coagulase test virtual interactive bacteriology laboratory. If possible, perform the test under a biosafety hood. Your instructor will direct you as to which procedure to use, but the tube test is the definitive test. If so what is the best and most cost effective procedure to perform the tube coagulase test. The bound coagulase is also known as clumping factor. This test demonstrate the presence of catalase, an enzyme that catalyses. Dnase test agar is not intended for use in the diagnosis of disease or other conditions in humans. The catalase test is used primarily in differentiation between certain genera and species of bacteria.
The test is used to differentiate staphylococcus aureus positive from coagulasenegative staphylococci negative. This enzymatic test is essential in the scheme of identification for grampositive organisms and certain gramnegative organisms. Coagulaseproduction staphylococcus differing traditional. Catalase test catalase test catalase is the enzyme that breaks hydrogen peroxide h 2o2 into h 2o and o 2. While there are seven antigenic types of free coagulase, only one antigenic type of bound coagulase exists. This video lesson demonstrates how to interpret the results of the coagulase test used in identifying staphylococcus aureus. Coagulase test slide for microbiology slide coagulase. The coagulase test can be performed using two different procedures slide test and tube test. Choose from 200 different sets of coagulase test flashcards on quizlet. Please update to the latest version of flash player. Slide coagulase test 1 remove a single tube of coagulase cryo from the freezer and permit it to thaw to room temperature. The coagulase test has traditionally been used to differentiate staphylococcus aureus from coagulasenegative staphylococci. The slide test is simple, giving results within 10 seconds, but it can give false negatives. Using a sterile loop emulsify 24 colonies of the test organism into the plasma.
To introduce and demonstrate the principle and experimental set up for determining the microbes ability to detoxify hydrogen peroxide and or to cause blood coagulation. The coagulase test is used to differentiate species of staphylococcus, especially the coagulase positive staphylococcus aureus from coagulase negative staphylococcal species. Coagulase test is carried out to differentiate potentially pathogenic staphylococcus species from other gram positive, catalasepositive cocci by the detection of coagulase. Bound coagulase, otherwise known as clumping factor, can be detected by carrying out a slide coagulase test, and free coagulase can be detected using a tube coagulase test. Other staphylococci do not produce coagulase, thus this test can distinguish s.
Laboratory procedure bbl staphyloslide latex test for staphylococcus aureus i. The tube test is the definitive test, however, it can take up to 24 hours to complete. Its concentration in bacteria has been correlated with pathogenicity. Slide coagulase test may be used to screen isolates of s. Free coagulase is detected in tube coagulase test and bound coagulase is detected in slide coagulase test. Uk standards for microbiology investigations smis are developed under the auspices of public health england phe working in partnership with the national health service nhs, public health wales and with the professional organisations. Pdf tube coagulase testing for the identification of. Product summary and explanation in 1956, weckman and catlin showed a correlation between increased dnase activity of staphylococcus aureus and positive coagulase activity. In addition, the microtube method offers a more convenient and economical format for the performance of the coagulase tube test. Catalase is the enzyme that catalyzes to split up hydrogen peroxide into its reactant water and oxygen. The catalase test is used to detect the presence of the enzyme catalase in bacteria. Emulsify several isolated colonies of test organism in 1 ml of diluted rabbit plasma to give a milky suspension.
Discussion when s aureus is suspected most hospital microbiologists usually perform a simple confirmatory test at the bench. Slide test to detect bound coagulase place a drop of physiological saline on each end of a slide, or on two separate slides. It is a waste to run the test on a nonstaph isolate. Field and smith 8 compared the plasmas of several animal species for use in the slide test. This version of the coagulase test is used to identify the presence of either bound coagulase or free coagulase, which is an extracellular enzyme. The coagulase tube test has been set up as a demo for you to observe in class. This protocol describes the history and procedures of the coagulase test. Full text get a printable copy pdf file of the complete article 663k, or click on a page image below to browse page by page. The tube coagulase test is thought to be the more definitive of the two, however, it can take several hours to overnight to produce a result. In biochemistry, tests tags catalase test, catalase test precautions, catalase test priniciple, catalase test procedure, results july 22, 2016 2587 views hamza khan. It is an important indicator for the pathogenicity of staphylococcus strains. It is concluded that in most instances, the latex slide agglutination test is areliable methodfor identifying s. The coagulase test is used to differentiate species of staphylococcus, especially the coagulasepositive staphylococcus aureus from coagulasenegative staphylococcal species. The widely promulgated interpretation that all degrees of clotting in coagulase plasma are a positive identification of s.
Coagulase slide tests have been carried out on food, human and animal strains of staphylococci with human and animal plasmas. This video was prepared by the teaching support team for the university of western australias school of pathology and laboratory medicine palm. The tube coagulase test detects both bound and free coagulase. The slide test is performed by preparing a suspension of bacterial cells mixed into a drop of rabbit plasma on a microscope slide. Catalase is an enzyme present in most cytochrome containing aerobic and facultative anaerobic bacteria. The latex test paralleled the tubetest butwithlowersensitivity. Learn coagulase test with free interactive flashcards. Microtube coagulase test for detection of coagulase. Both common versions of the test, the slide method and the test tube method, are described, and the mechanisms of the reactions are discussed. Coagulase test the enzyme coagulase, produced by a few of the staphylococcus species, is a key. Provide alternate content for browsers that do not support scripting or for those that have scripting disabled. Indeed, according to the manufacturers instructions, s intermedius strains would be identified as coagulase negative.
Numerous plating media have been preparedwithhumanorrabbit plasmas 6, 7, 10, 15, 17. Bound coagulase adsorbs fibrinogen from the plasma and alters it so it precipitates on the staphylococci, causing them to clump resulting in cell agglutination. Using a sterile 1 ml serological pipette, add approximately 0. Rapidly identify and differentiate staphylococci which possess coagulase andor protein a, particularly staphylococcus aureus, using the thermo scientific staphaurex latex agglutination test. In addition to potential selfcontamination by aerosol exposure, mycobacteria speciation by the catalase test is achieved by different techniques not discussed in this protoc ol 3, 5, 7. The tube coagulase test is a valid means of identifying staphylococcus aureus, provided that only a firm clot that does not move when the tube is tipped is considered a positive reaction. On isolates from critical specimens, a negative coagulase test obtained with an inoculum of less than two colonies should be confirmed by repeating the procedure with a larger inoculum or by using.