mb rec exam # 5

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  1. What kind of organisms are fungi?
  2. If fungi is single-celled it is a
  3. If fungi is filamentous it is a
  4. Candida A microscopic morphology
  5. Candida A gram stain reaction
    Gram +
  6. Candida A on Sabouraud Dextrose Acid Agar
    Moist, white, opaque
  7. Candida A in sabourauds dextrose acid broth
    clear with sediment "+"
  8. Candida A chlmydocondia test
    + (terminal clomisdospore)
  9. Candida A species ID
  10. Candida B microscope morphology
    Enlongated cigar shaped
  11. Candida B gram stain
    Gram +
  12. Candida B on sabouraud dextrose acid agar
    dry, off-white, translucent
  13. Candida B growth in sabouraud's dextrose acid broth
    flaky, white surface growth
  14. Candida B chlamydoconidia test
    "-" (cross sticks/pseudohyphae)
  15. Candida B species ID
  16. What are the two media used to id dermatophytes?
    • dermatophyte test medium (DTM)
    • Sabouraud dextrose acid agar
  17. What are you looking for in the DTM? (Dermatophyte Test Medium)
    Production of red color
  18. what are you looking for in sabouraud Dextrose Acid Agar?
    observation of macroscopic appearnace of growth
  19. Microsporum audouinii appearance
    white cottony colony
  20. Microsporum audouinii is the causative agent of ?
    tinea capitis
  21. Trichophyton rubrum appearance?
    white cottony colony, red on reverse side
  22. Trichophyton rubrumis the causative agent of?
    tinea pedis
  23. Trichophyton tonsurans appearance?
    colonies from a folded surface with yellowish color in the depressions. Surface is velevety to powdery with a variation in color.
  24. Trichophyton tonsurans is the causative agent of?
    tinea capitis
  25. is cryptococcus neoformans yeast or mold?
    always yeast
  26. What is needed to identify the capsule of a yeast?
    india ink stain
  27. Why is it hard to stain the capsules of the yeast?
    very mucoid colony ( large amount of polysaccharide)
  28. what test is used to id the genus of yeast?
    Urease Test
  29. Cryptococcus sp. urease test result?
    positive (pink color)
  30. Candida sp. urease test result?
    Negative (No color change)
  31. Yeast cells other than _______ are urease negative
  32. cryptococcus sp is a yeast or mold?
  33. candida sp is a yeast or mold?
  34. What is used to id species in yeast?
    Nigerseed (birdseed) agar
  35. cryptococcus neoformans nigerseed agar result?
    brown colored colonies
  36. cryptococcus sp. (other than cryptococcus neoformans) nigerseed agar result?
    White colonies
  37. What is dimorphic fungi at 25 C?
  38. What is dimorphic fungi at 37 C?
  39. What is sporothrix schenckii?
    a mold colony that has characteristic mitocondria in daisy cluster
  40. Mycobacteria
    aerobic slow growing bacilli that possess a high lipid content in their cell wall
  41. What oganisms are "acid-fast bacilli"
  42. what is required for the decolorization of mycobacteria?
    a spcial stain that utilizes acid alcohol
  43. What are the four special chemicals used to stain mycobacteria?
    • carbol fuschin solution (kinyoun) containing tergitol
    • acid alcohol solution
    • methylene blue, alkaline
    • broth (Dubos) culture of mycobacterium grodonae
  44. list the 11 steps of mycobacteria staining
    • prepare films of resuspended broth culture and fix slide by gently heating
    • place slide on slide rack
    • cover film with carbol fuchsin (kinyoun)
    • allow to stand for 5 minutes
    • wash off with water
    • wash with acid alcohol until decolorized (grayish appearance)
    • wash off with water
    • counterstain with methylene blue, alkaline for (15-30 secs)
    • wash off with water
    • dry by blotting
    • examine under oil immersion
  45. when staining mycobacteria what appears red?
    acid-fast bacteria
  46. when staining mycobacteria what stains blue?
    bacteria which are not acid-fast stain blue
  47. What part of the body is needed to stain for mycobacteria?
  48. rice extract agar is a suitable substrate for what?
    chlamydospore production
  49. When candida albicans is grown on rice extract agar, what happens?
    mycelium-bearing ball-like clusters of budding cells and the thick-walled round chlamydospores
  50. What is the general process of the germ tube testing?
    • Take yeast cells and serum
    • incubate it for (2-4 hours)
    • wait for germ tube formations
  51. DTM is selective for the growth of what?
  52. Why is the ph indicator (phenol red) added to DTM?
    To monitor acidic/basic reactions
  53. A yellow color in DTM means what?
    reaction is acidic
  54. A red color on the DTM means what?
    Reaction is basic
  55. What happens on the DTM to make its pH rise and turn it red?
    dermatophytes utilize the keratin in the medium they produce alkaline end products.
  56. what is the disease name of sporothrix schenckii?
    "Rose Gardner's disease"
  57. how do you concentrate sputum for culture, film and antibiotic observations (mycobacterium)
    • Get a sputum specimen of at least 10ml
    • add 10ml of digestant = (NaOH/NALC)
    • vortex 30-60 seconds (liquified)
    • let stand at R.T for 15-20 minutes
    • add 30ml phosphate buffer (pH = 6.8)
    • centrifuge at 3,000 rpm for 30 minutes
    • discard supernatant
    • take the sputum sediment ( for antibiotic sediment,for culture = lowenstein Jensen & Middlebrook 7H-10/7H-11, for film= acid fast stain )
  58. viruses are obligate ________ _________
    intracellular parasites
  59. what do viruses require for replication?
    Host cells
  60. can be viruses be grown or cultured in artficial media?
    no, only bacteria and fungi
  61. what are the three nonimmunological methods to IDing viruses?
    • electron microscopy
    • virus growth
    • detection of viral genome
  62. what virus is readily diagnosed by electron microscopy of stool samples?
  63. what is the causative agent of infantile gastroenteritis ?
  64. Whtat can viruses be grown in?
    • tissue culture
    • embroyonated chicken eggs
    • experimental animals
  65. What media is the prime choice for growing and isolation of the influenza viruses?
    embryonated chicken eggs
  66. where are most viruses grown?
    Tissue cultures
  67. What is the ID of a virus based on?
    the CPE followed by id of virus specific antigens by immunoflorescence of other immunological technique
  68. CPE
    degenerative changes in cells, especially in tissue culture, and may be associated with the multiplication of certain viruses.
  69. what is used to detect viral genome during viremia?
    (PCR & RT-PCR) - Polymerase chain reaction and reverse transcriptase
  70. (PCR & RT-PCR) can be used to id what
    DNA or RNA
  71. Immunologic methods are used to what?
    ID viruses and quantify the immune response(antibodies) in response to the etiologic (causative) agent during the acute and convalescent stages of the illness.
  72. what are the immunological methods that can be used to id viruses?
    • immunofluorescence
    • enzyme immunoassay (EIA)
    • enzyme-linked immunosorbebt Assay (ELISA)
    • western blot analysis
    • hemagglutination- inhibition test
    • neutralization of viral infectivity
  73. what does NALC stand for?
  74. In the concentrated sputum process what is NALC?
    digestant (mucolytic)
  75. In the concentrated sputum process what is NAOH?
  76. what inhibits the growth of microorganism other than mycobacteria on the loenstein jensen culture media?
    additive malachite green
  77. what is the medium of choice to test mycobacterium antibiotic susceptibility?
    middlebrook 7h-10/7h-11
  78. mycobacterium staining process called?
    Ziehl-Neelsen (ZN) method
  79. total magnification for electron microscope?
  80. two types of immunoflourences?
    indirect and direct
  81. direct IF?
    when a fluorescence molecule is covalently linked to the antibody molecule
  82. indirect IF?
    in addition to first antiobdy in the direct version, a fluroescent second antibody specific for the primary antibody,
  83. EIA (enzyme immunoassay)
    An enzyme like horseradish peroxidase or alkaline phosphatase is conjugated to the antibody after the antibody-antigen reaction, enzyme substrate is added which converts the substrate into a chromophore at the site of the antigen-antibody complex.
  84. what does ELISA stand for?
    Enzyme-linked-immunosorbent assay
  85. ELISA positive reaction steps?
    • 1. antibody present in serum
    • 2. binding of secondary conjugated antibody
    • 3. enzyme reacts with substrate
  86. ELISA Negative reaction steps?
    • 1. antibody absent from serum
    • 2. conjugated antibody does not bind
    • 3. no conversion of substrate
  87. ELISA is used for screening for what?
    HIV antibodies
  88. The darker the color for ELISA means ...?
    the more Antibodies there is
  89. in ELISA color means
    antibodies (usually yellow)
  90. what kind of test is the western blot analysis?
  91. in the western blot anaylisis, viral proteins are seperated by what?
    polyacrylamide electrophoresis
  92. in the western blot anaylsis, the protein bands are transferred to where?
    nitrocellulose paper
  93. viral proteins in the western blot analysis are .?
  94. what is added to the nitrocellulose paper to react with the protein bands?
    enzyme-conjugated anti-immunoglobulin reagent
  95. What type of test is the oraquick rapid HIV?
  96. What samples can be used for the oraquick rapid HIV test?
    blood, plasma or oral fluids
  97. what is the confirmatory test that should be used with the oraquick rapid hiv test?
    western blot analysis
  98. The virus hemagglutination (HA) titer indicates that how many HA units be used for the hemagglutination-Inhibition (HI)
    4 HA units
  99. How do you calculate the dilution at which the tests are obtained?
    divide the HA titer by 4
  100. how do you calculate the fold increase in the acute and covalescent titers?
    divide the covalescent titer by the acute titer.
  101. What is the main point/process of the neutralization of viral infectivity
    neutralization of viral CPE by antibodies.
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mb rec exam # 5
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