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    Instrucciones Coagulasa

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    shealock
    The document describes a test for detecting the coagulase enzyme produced by Staphylococcus aureus. It explains that coagulase causes plasma to coagulate. The test involves incubating bacterial cultures with rabbit plasma and observing for coagulation, with varying degrees of coagulation indicating different levels of coagulase production.

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd

    Instrucciones Coagulasa

    Uploaded by

    shealock
    256 views1 page
    The document describes a test for detecting the coagulase enzyme produced by Staphylococcus aureus. It explains that coagulase causes plasma to coagulate. The test involves incubating bacterial cultures with rabbit plasma and observing for coagulation, with varying degrees of coagulation indicating different levels of coagulase production.

    Original Description:

    Instrucciones Bactident Coagulasa

    Original Title

    Instrucciones coagulasa

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    © © All Rights Reserved

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    The document describes a test for detecting the coagulase enzyme produced by Staphylococcus aureus. It explains that coagulase causes plasma to coagulate. The test involves incubating bacterial cultures with rabbit plasma and observing for coagulation, with varying degrees of coagulation indicating different levels of coagulase production.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    256 views1 page

    Instrucciones Coagulasa

    Uploaded by

    shealock
    The document describes a test for detecting the coagulase enzyme produced by Staphylococcus aureus. It explains that coagulase causes plasma to coagulate. The test involves incubating bacterial cultures with rabbit plasma and observing for coagulation, with varying degrees of coagulation indicating different levels of coagulase production.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 1

    Bactident Coagulase

    For the detection of the enzyme coagulase developed by staphylococcus aureus.


    Bactident Coagulase

    Mode of Action
    Coagulase is an enzyme with the ability to coagulate plasma.
    Staphylococcus aureus forms two types of coagulase. The free
    coagulase is an extracellular enzyme, the bound coagulase is
    localized on the surface of the cell wall. Both enzymes are
    detected in the tube test. With the slide test, only the bound
    coagulase can be detected.

    If the test is negative after 4-6 hours, continue incubating the


    tube and make a final assessment after 24hours. For the
    negative control, prepare a brain-heart broth, but do not
    inoculate. There must be no sign of clotting.
    For the positive control, conduct the test with a coagulasepositive strain of staphylococcus.

    Method

    negative

    no coagulation

    An overnight broth culture of staph. aureus is incubated with


    rehydrated EDTA-rabbit plasma. The coagulase test is positive, if
    more than three quarters of the tube contents forms a coherent
    clot.

    1 + positive

    a few small separate clots

    2 + positive

    a few small joined clots

    3 + positive

    large extensively coagulated clots

    Stability

    4 + positive

    complete coagulation, contents do not shift


    when tube is inverted

    See expiry date.


    Bactident Coagulase is stable for 5 days in the dissolved
    (rehydrated) condition at +2C to +8C. At -20C it is stable for
    up to 30 days.

    Storage
    Store cool, dry and tightly closed at +2C to +8C. Store
    dissolved plasma at +2C to +8 C or deepfrozen at -20C. Do
    not refreeze once defrozen.

    Notes
    The slide test in which a colony is mixed with rabbit plasma on
    a microscope slide (clumping factor), only detects the bound
    coagulase and can at best only service as a screening test. Falsepositive reactions and autoagglutination can occur.

    Ordering Information

    Safe Disposal

    Product

    Merck Cat. No.

    The contents of the tube include bacteria and must be disposed


    of safely. This can be done by autoclaving or placement in a
    5-6% solution of disinfectant for at least 6hours.

    Pack
    contents

    Bactident Coagulase

    1.13306.0001

    6 vials each
    containing
    3ml of
    lyophilized
    rabbit plasma
    with EDTA

    Experimental Procedure
    a. Conduct the coagulase test on 5 typical and/or 5 a-typical
    colonies on BAIRD-PARKER agar (Merck Cat. No. 1.05406.)
    or 5 suspect colonies from other culture media (CHAPMAN
    agar, Merck Cat. No. 1.05469., VOGEL-JOHNSON agar,
    Merck Cat. No. 1.05405., Blood agar (base), Merck Cat. No.
    1.10886.).
    b. Transfer each of the selected colonies with a sterile
    inoculation loop to separate culture tubes containing brainheart broth (Merck Cat. No. 1.10493.) and incubate at 37C
    for 20-24 hours.
    c. Dissolve the freeze-dried EDTA-rabbit plasma in 3ml of
    distilled or demineralized water.
    d. Pipette 0.3ml of the rehydrated Bactident Coagulase into a
    sterile culture tube using a sterile pipette.
    e. Carefully mix 0.1ml of the brain-heart broth culture 1/2 an
    inoculation loop of colony material from the CHAPMAN
    blood or BAIRD-PARKER agar with the 0.3ml of plasma and
    incubate in a water bath at 37 C. (Colony material directly
    from VOGEL-JOHNSON or mannitol-sodium chloride-phenol red agar is not suitable for the test. A brain-heart broth
    culture is required first.).
    f. Every hour, check the tube contents for coagulation by
    gently tipping to the side (not by shaking).
    g. The coagulase test is positive, if more than three quarters of
    the tube contents has formed a coherent clot.
    Merck Microbiology Manual 12th Edition

    179

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