Serological Procedures 6.2

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Serological Procedures 6.2
2010-08-17 15:02:12
Serology serological procedures NSHS MLT

serological procedures 6.2
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  1. The ability of a particular antibody to combine with one antigen instead of another.
  2. fit between the antigenic determinant and the combining site of the antibody.
    non-covalent force
  3. some of the determinants of an antigen are shared by apparently unrelated molecules.
  4. strength of a multivalent antibody to bind to a multivalent antigen
  5. the bond between a single antigenic determinant and an individual combining site of an antibody.
  6. excessive amount of antibody that can give a false negative result.
  7. decreased amount of antibodyt caused by an excess of antigen
  8. point of equal antigen and antibody amounts which yield the greatest reaction.
    zone of equivalence
  9. deviance from the optimum pH of ___ will alter reactivity of the antibody.
  10. different classes of antibodies react at different __________.
  11. what is the antibody that reacts at warm temperatures?
  12. what is the antibody that reacts at cold temperatures?
  13. incubatin time varies with the _____ of antibody.
  14. which types of antibodies will be all the same class and have only one type of heavy and light chain?
    monoclonal antibodies
  15. these are uniform, highly specific antibodies that caon be produced in large quantitites, cloned from a single cell to bind to a single specific antigen.
    Monoclonal antibodies
  16. what are the three uses of monoclonal antibodies?
    • typing erythrocyte antigens
    • typing leukocyte antigens
    • as a reagent
  17. serological samples should be centrifuged for __ minutes at moderate speed.
  18. if testing cannot be done promptly the serum should be frozen at what temperature?
  19. how is complement inactivated?
    heat serum to 56oC for 30min
  20. if serum is not used within four hours, reheat serum at 56oC for ___ minutes.
  21. the highest dilution in which a reaction occurs.
  22. what is the purpose of a titer?
    to determine the concentration of antibody
  23. this is used to make serum less concentrated.
  24. each succeeding dilution is the same as the preceding dilution
  25. calculations of any given dilution can be determined by using this formula:
  26. in the formula D1(V1)=D2(V2) what is the starting dilution.
  27. in the formula D1(V1)=D2(V2) what is the starting volume?
  28. in the formula D1(V1)=D2(V2) what is the final dilution
  29. in the formula D1(V1)=D2(V2) what is the final total volume?
  30. pipettes should be periodically calibrated for what three things?
    • accuracy
    • precision
    • reproducibility
  31. Red Blood cell Suspensions should be used in serological tests as an ________.
  32. in an RBC suspension what should be used as the diluent?
    2-5% NaCl
  33. RBC's contain _______; serve as receptor sites for antibodies or complement.
  34. what are the four carriers used in serology?
    • carbon
    • RBC's
    • Latex
    • Bacteria
  35. what is the calucation that can be used to determine the minimum packed cell volume needed to make a solution?
    PCV= (total volume X % desired)/100
  36. in an RBC suspension this represents the packed RBCs.
  37. true or false
    Hemolyzed RBC's can be used in an RBC suspension.
  38. Mono and ASO use what carrier?
  39. Floculation tests use what carrier?
  40. what carrier is used in hemaglutination?
  41. in what testing method, if antigens are present in serum, they will bind with antibodies forming visible cross-linked aggregates.
    latex agglutination
  42. in latex agglutination antibody molecules bind to latex _____.
  43. what are the five latex agglutination immunologic assays?
    • antistreptolysin O
    • Mono Test
    • C-reactive protein
    • IgG and IgM rheumatoid factors
    • Rubella antibody
  44. in what test are latex particles coated with strptolysin O antigen?
    Antistreptolysin O
  45. in Antistreptolysin O visible agglutination will be seen mixed with patient's serum containing what?
    ASO antibody
  46. titers begin to rise at __ days after the onset of infection and peaks at ____ weeks after infection.
    • 7 days
    • 3-6 weeks
  47. In ASO, a four fold increase between acute and convalescent tube indicates a recent infection with what?
    Group A strep
  48. ASO antibodies are elevated in acute/convalescent sera with what disease?
    acute rheumatic fever
  49. serological testing should compare acute and convalescent sera collected ______ weeks apart.
  50. how long are ASO samples stable at 2-8oC?
    eight days
  51. how long are ASO specimens stable at -25oC?
    3 months
  52. this is based on agglutination of horse RBC's by hetrophil antibody present in infectious mononucleosis.
  53. in monotest horse RBC's exhibit antigens against both __________ and ______________ threfore a differential absorption of patient serum is necessary.
    • forssman
    • infectious mononucleosis
  54. the monotest immunologic theory is based on __________ principle.
    Davidshon principle
  55. in the monotest immunologic theory, what contains only forssman antigen which will absorb forssman heterophil antibody?
    guinea pig kidney
  56. in monotest immunologic theory what will absorb only no-forssman heterophil antibody of infectious mononucleosis?
    beef RBC's
  57. in a monotest immunologic theory this is tested with both the guniea pig-serum specimen and the beef RBC-serum specimen.
    horse RBC's
  58. to be positive for IM heterophil antibody, the guinea pig should be _________ for agglutination and the beef RBC should be _________ for agglutination.
    • Guinea pig: positive
    • Beef RBC: negative
  59. what is the sample for Monotest?
    Serum or plasma mixed with EDTA
  60. what is the confirmatory test for infectious mononucleosis?
    Epstein-Barr virus antibody
  61. this is the causitive agent of infectious mononucleosis?
    ebstein-barr virus
  62. this is a DNA herpes-type virus that infects the B-lymphocytes?
  63. this is tested to confirm infectious mononucleosis on immune suppressed patients.
    EBV antibodies
  64. this test is beneficial in defining immune status and stage of infectious mononucleosis.
  65. antigen-antibody __________ when combined in proportions at or near equivalence.
  66. in what type of test is interaction of soluble antigen with antibody that results in the formation of a precipitate of fine particles?
  67. in flocculation test particles are ____________ or _____________ visible.
    • macroscopically
    • microscopically
  68. what are two examples of flocculation tests?
    • VDRL
    • RPR
  69. this test is based on agglutination of RBC's with antigens that are natural or bound to RBC's
  70. what are three examples of hemagglutination immunologic assays?
    • ABO antibodies
    • Rh antigens
    • Cold Agglutinins
  71. Precipitation methods are performed in what two mediums?
    • semi solid media (agar or agarose)
    • non-gel support medium (cellulose acetate)
  72. what are the two types of precipitation methods?
    • double immunodifusion
    • radial immunodifusion
  73. in this test antibody dilutions and specific antigens are placed in adjacent wells. Antigen and antibody diffuse out, bind together to form a visible precipitate.
    Double immunodiffusion
  74. what are the three basic reaction patterns in double immunodiffusion?
    • Identity
    • Non-identity
    • partial identity
  75. In which double immunodiffusion result in precipitin band forms a single smooth arch?
  76. in which double immunodiffusion resuld do precipitin lines merge in a spur formation?
    partial identity
  77. in which double immunodiffusion reslut do precipitin lines cross each other?
  78. what are the acceptable specimens for double immunodiffusion?
    • serum
    • urine
  79. in double immunodiffusion why are urine specimens tested in concentrated and unconcentrated forms?
    wide range of light cahin concentrations
  80. what are the two double immunodiffusion immunologic assays?
    • Rheumatoid arthritis
    • systemic lupus erythrematosus
  81. Radial immunodiffusion (RID) is used to identify and quantitate the number of _________ found in human serum.
  82. in radial immuno diffusion (RID) the antigen will diffuse throughout the agan and form what?
    a precipitin ring
  83. in radial immunodiffusion specific antibody is added to a buffered agarose medium, test ________ is added to a well in the agarose.
  84. in radial immunodiffusion (RID) diameter of the precipitin rig is related to what?
    antigen concentration
  85. what are the four clinical applications of radial immunodiffusion (RID)?
    • Alpha-1-antitrypsin
    • Transferrin
    • C3
    • Immunoglobulin
  86. what are the two electro immunodiffusion methods?
    • Countercurrent immunoelectrophoresis (CIE)
    • rocket immunoelectrophoresis
  87. in what test do antigens and antibodies move more quickly toward each other with addition of electrical current?
    countercurrent immunoelectrophoresis (CIE)
  88. in CIE agarose gel is used. pH is such that antibodies are ________ charged and antigens are ________ charged.
    • antibodies: positively
    • antigen: negatively
  89. in countercurrent immiunoelectrophoresis how is the zone of equivalence represented?
    antigen-antibody complex forsm a visible line of precipitin
  90. what is the clinical application of countercurrent immunoelectrophoresis (CIE)?
    • detection of anti-nuclear ribonucleoprotein (nRNP):
    • associated to sharp's syndrome (mixed connective tissue disease)
  91. what test is associated with sharp's syndrome (mixed conective tissue disease)?
    detection of anti-nuclear ribonucleoprotein (nRNP)
  92. in rocket electrophoresis is based on antigen ______________ in an antibody containing gel.
  93. in rocket electrophoresis what is adjusted to inhibit antibody migration?
  94. in rocket electrophoresis the height of the rocket is __________ to the antigen concentration.
  95. what ar the four types of labeling techniques?
    • radio immuno assay (RIA)
    • chemiluminescence
    • enzyme immunoassay (EIA)
    • immunofluorescent techniques
  96. what are the two assays that involve chemiluminescence?
    • competitive assay
    • sandwich assay
  97. what are the two types of immunofluorescent techniques?
    • direct
    • indirect
  98. in what test are radioisotopes used to measure the concentration of antigens or antibodies
    radioimmunoassay (RIA)
  99. in what test are radiolabled ligands bound to antigen-antibody complex?
  100. in RIA radiactivity is counted on what?
    gamma counter
  101. this test is extremly sensitive and able to detect trace amounts of antigen or antibody.
    radioimmunoassay (RIA)
  102. what are the two clinical applications of radioimmunoassay (RIA)
    • hepatitis A IgM antibody
    • hepatitis A antigen
  103. in this test lables can be attached to antigen or antibody.
  104. in this chemiluminescence assay, a fixed amount of labeled antigen competes with unlabeled antigen (patient sample) for a limited numberr of antibody-binding sites.
    competitive assay
  105. in a competitive assay the amount of light emitted is _________ proportional to the amount of analyte (antigen) being tested.
  106. in this chemiluminescene test sample antigen binds to a fixed antibody on a solid phase, then a second antibody labled with chemiluminescent binds to the antigen-Ab complex on the solid phase.
    sandwich assay
  107. in a sadwich assay the amount of light emitted is __________ proportional to the amount of the analyte (antigen) being tested.
  108. what are the two clinical applications of chemiluminescence?
    • hepatitis A IgM antibody
    • hepatitis A antigen
  109. this test is similar to RIA but used a non-isotopic label.
    enzyme immunoassay
  110. this test is safer than RIA yet has the same specificity and sensitivity as RIA.
    enzyme immunoassay (EIA)
  111. in Enzyme immunoassay (EIA) what is used instead of a radioisotope?
    an enzyme labeled antibody or antigen conjugate
  112. what ar the most commonly used enzymes to detect the presence and quantity of antigen or antibody in patient serum?
    • peroxidase
    • alkaline phosphatase
  113. in EIA substrate causes color production and is measured how?
  114. what are the three clinical applications of Enzyme Immunoassay (EIA)?
    • hepatitis B
    • HIV antibody
    • Cytomegalovirus antigen/antibody
  115. in this extremely specific and sensitive test, fluorescent molecules are used instead of radioisotopes or enzyme labels.
    immunofluorescent techniques
  116. what are the three clinical applications of immunofluorescent techniques?
    • CMV antigen
    • CMV antibodies
    • HIV antibodies
  117. in this immunofluorescene technique conjugated reagent antibody is used to detect antigen-antibody complex at microscope level.
    Direct Immunofluorescence
  118. in direct immunofluorescence what is used ot visualized many bacteria in direct smears?
    fuorescein-conjugated antibodies boud to fuorochrome
  119. direct immunofluorescence uses the immunological/physiological theory of:
    intracellular antigen detection
  120. what are the two sepcimens used for direct immunofluorescence?
    • tissue
    • bacteria smears
  121. in this test, antigen source such as toxoplasma microorganism or virus of infected tissue culture cells to the specific antibody being tested is affixed to a slide.
    indirect immunofluorescence
  122. in indirect immunofluorescence what reacts with antigen forming an antigen-antibody complex?
    unkown antibody (patient's Ab)
  123. in indirect immunofluorescence what is added to the Ab-Ag complex to bind to the antibody as a marker?
    labeled anti-immunoglobulin (fluorescent Ab)
  124. indirect immunofluorescence measures what?
  125. direct immunofluorescene measures what?
  126. what is the specimen collected for indirect immunofluorescence?
  127. what are three indications for performing indirect immunofluorescene on serum?
    • toxoplasma microorganism
    • virus infected tissue culture cells
    • ANA antibodies
  128. what is the classic method for demonstrating presence of a complement fixing antibody in serum?
    complement fixation
  129. what is the indicator system for complement fixation?
    • sheep RBC's
    • anti-sheep antibody
    • complement from guinea pig serum
  130. in complement fixation patient serum is added to _______ antigen, and complement is added to the solution.
  131. in complement fixation, if serum contains the antibody for the antigen, then an antigen-antibody complex will bind with what?
  132. in complement fixation sheep RBC's with anti-sheep antibody are added to the solution of antibodies, antigen, and complement. if complement has not been bound, it is available to bind with what?
    coated sheep RBC's
  133. hemolysis of indicator sheep cells indicates a lack of antibody in patient serum and a _________ complement fixation test.
  134. how is a positive complement fixation test demonstrated.
    lack of hemolysis
  135. why is a positive complement fixation test demonstrated by the lack of hemolysis in indicator sheeps RBC's?
    patients serum does contain a complement fixing antibody
  136. what are the three clinical applications of complement fixation?
    • Varicella-zoster virus
    • Rubella virus
    • Herpes simplex infection
  137. this is an in vitro method that amplifies low level of specific DNA sequences in a sample to higher quantities.
    polymerase Chain Reaction
  138. in PCR ___ short DNA primers are used as templates that follow the enzymatic process.
  139. in PCR the enzymatic prcess is carried out in cycles and repeated up to ___ times.
  140. what are the four steps of enzymatic process in PCR?
    • DNA denaturing
    • Primer anealing
    • Extension of primed DNA sequenced
    • The amount specific DNA sequence double and examined in various ways
  141. polymerase chain reaction is used in the detection of ____.
  142. what are the two clinical applications for PCR?
    • human papilloman virus (HPV)
    • coronary artery disease
  143. If the nucleic acid of interest is RNA rather than DNA, the PCR procedures can be modified to include the conversion of RNA to DNA using what?
    Reverse Transcriptase-Polymerase (RT-PCR)
  144. the clinical application for RT-PCR is to test for what?
  145. what are the two clinical applications of RNA?
    • Human Immunodeficiency Virus
    • Hepatitis C Virus