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219 Cards in this Set
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Initial force of Attraction that exists between a single FAB site on an Antibody and single epitope or determinant site on the corresponding antigen |
Affinity |
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Antigen-Antibody Binding is held together by |
Weak Non-covalent bonds |
Irreversible |
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Non-covalent Bonds: |
1. Ionic Bond 2. Hydrogen Bond 3. Hydrophobic Bonds 4. Van der Waals |
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Sum of all the attractive forces between antigen and Antibody Measure of overall stability of an antigen-Antibody complex |
Avidity |
Strength with which a multivalent Antibody binds a multivalent antigen |
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Caused by presence of antigenic determinants that resemble one another Can be avoided by using Monoclonal Antibodies |
Cross Reactivity |
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Agglutination tests are ___________________ tests only |
Screening |
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Physical conditions purposely manipulated to break the antigen-Antibody complex, with subsequent release of the Antibody into the surrounding medium |
Elution |
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Antigenic Determinant |
Epitope |
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Part of Antibody that binds to Antigen |
Paratope |
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Causes of False Positive reactions in Agglutination Testing |
1. Overcentri 2. Contaminated glasswares, slides, reagents 3. Autoagglutination 4. Saline stored in Glass bottles 5. Cross-Reactivity 6. RF, Heterophile Ab 7. Delay in reading the slide
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Causes of False Negative reactions in Agglutination Testing |
1. Undercentri 2. Inadequate washing of cells 3. Reagents not active 4. Delays in Testing especially AHG 5. Incorrect incubation Temp 6. Insufficient incubation Time 7. Prozone 8. Failure to Add AHG reagent |
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False appearance of clumping which may be due to rouleaux formation |
Pseudoagglutination |
Maybe dispersed by adding a few drops of PHYSIOLOGIC NACL |
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Causes of Rouleaux Formation |
1. High or Abnormal Globulins 2. Plasma Expanders (Dextran) |
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Involves combination of soluble antigen with soluble Antibody to produce insoluble complexes |
Precipitation |
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Where optimum precipitation occurs Number of multivalent sites of antigen and Antibody are approximately equal |
Zone of Equivalence |
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Excess ANTIBODY |
PROZONE |
False Negative Remedy: Serum Dilution |
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Excess ANTIGEN |
POSTZONE |
False Neg Remedy: Repeat test after a week |
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Measures amount of light blocked |
Turbidimetry |
Light detection is in direct line with the incident light |
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Measures the amount of light scattered |
Nephelometry |
Light detection device is in angle from incident light |
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Antigen and Antibody diffuse through agar gel and precipitate when they reach zone of equivalence No electric current used to speed up the process |
Passive Immunodifusion |
Support Medium: 1. Gel 2. Agar 3. Agarose |
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Precipitation by Passive Immunodiffusion |
1. Single Linear Immunodiffusion 2. Single Radial Immunodiffusion - Mancini/Endpoint Method - Fahey and McKelvey/Kinetic Method 3. Double Radial Immunodiffusion (Ouchterlony) |
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Single Linear Immunodiffusion Procedure |
1. Antibody is incorporated into agarose in a testube
2. Antigen is layered on top, moves down the gel and precipitation occurs |
Precipitation moves down the tube in proportion to antigen concentration |
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Single Radial Immunodiffusion Procedure |
1. Antibody is uniformly distributed in the support gel
2. Antigen is placed in a well cut into the gel
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AREA OF RING obtained is a measure of antigen concentration |
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Antigen is allowed to diffuse to completion and when equivalence is reached, no further change in diameter
Square of Diameter is proportional to Antigen Concentration |
Mancini/Endpoint Method |
IgG 24 hrs IgM 50-72 hrs |
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Measurements are taken before point of equivalence is Reached
Stevens: 18 Hours Calbreath: 24 Hours Diameter is proportional to the log of antigen concentration |
Fahey and MacKelvey/Kinetic Method |
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Both antigen and Antibody diffuse independently through a semi-solid medium in two dimensions |
Double Radial Immunodiffusion (Ouchterlony) |
1. Antibody placed in Central Well 2. Antigens are placed in the surrounding Wells to determine if antigens share identical Epitopes |
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Double Radial Immunodiffusion (Ouchterlony) : Positive Result |
Precipitin Lines |
Serological Identity: SMOOTH ARC
Partial Identity: SPUR
Non-Identity: CROSSED LINES |
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RID + Electrophoresis Positive: Precipitin Rocket |
Rocket Immunoelectrophoresis/One-Dimension Electroimmunodiffusion |
1. Antibody is uniformly distributed in the gel
2. Antigen is placed in Wells cut in the gel |
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Serum (containing antigens) are electrophoresed
Antibody is placed in a Trough Positive: Precipitin Arc |
Immunoelectrophoresis |
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Serum (containing antigens) is electrophoresed
Antibody is overlaid directly to the gel's surface Positive: Precipitin Band |
Immunofixation Electrophoresis |
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Process by which particulate antigens such as cells aggregate to form larger complexes when a specific Antibody is present |
Agglutination |
1. Diret 2. Passive/Indirect 3. Reverse Passive 4. Agglutination Inhibition 5. Coagglutination 6. Antiglobulin-mediated |
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Antigen-Antibody combination through single antigenic determinant on the particle surface |
Sensitization |
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Sum of interactions between Antibody and Multiple Antigenic Determinants |
Lattice Formation |
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Antigen is found naturally on the surface of the particle |
Direct Agglutination |
Hemmaglutination Widal Test |
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Antigen is artificially attached to a carrier particle
Agglutination occurs if patient Antibody is present |
Passive/Indirect Agglutination |
Carrier Particles: 1. Bentonite 2. Charcoal 3. RBC 4. Latex 5. Gelatin 6. Silicates |
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Antibody is artificially attached to a carrier particle
Agglutination occurs if patient antigen is present |
Reverse Passive Agglutination |
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Competition between particulate and soluble antigens for limited Antibody-Binding sites
Positive: Lack Of Agglutination |
Agglutination Inhibition |
1. hCG 2. Hemagglutination Inhibition |
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Uses bacteria as inert Particles to which Antibody is attached S. aureus most frequently used because of its Protein A (naturally absorbs FC portion of IgG 1, 2, 4) |
Coagglutination |
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Detects nonagglutinating Antibody by means of coupling with a second Antibody (antihuman globulin) |
Antiglobulin- mediated Agglutination |
1. Direct Antiglobulin Test 2. Indirect Antiglobulin Test |
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In Vivo Sensitization |
Direct Antiglobulin Test |
Specimen: RED CELL Investigation of: 1. HDN 2. HTR 3. AIHA 4. DIHA |
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In Vitro sensitization |
Indirect Antiglobulin Test |
Specimen: Serum
Crossmatching Specimen: Serum
Ab determination and ID Specimen: Serum
Red Cell antigen phenotyping specimen: Patient RBC |
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Contains anti-IgG and anti-C3d obtained from Rabbits |
Polyspecific AHG |
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Contains anti-IgG or anti-C3d obtained from mice |
Monospecific AHG |
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Group O RBCs sensitized with IgG used to validate negative reaction |
O CHECK CELLS |
After addition of O check cells, Agglutination indicates: 1. AHG was added 2. AHG was not neutralized
Lack of Agglutination invalidates the results |
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Best at complement Fixation but fails to attach to Protein A |
IgG3 |
Protein A naturally adsorbs FC Portion of IgG 1, 2, 4 |
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Drugs causing positive DAT |
Drug adsorption: Penicillin
Membrane Modification: Cephalosporin
Immune Complex Formation: Rifampin, Phenacetin, Stibufen
Autoantibody Formation: Methyldopa, Mefenamic Acid |
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False Positive hCG testing |
1. hCG injection (pregnyl) 2. Chorioepithelioma 3. Hydatidiform mole 4. Excessive ingestion of aspirin |
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Measurement of the number of residual non-agglutinating Particles in a specimen using a laser beam in an optical particle counter |
Particle Counting Immunoassay (PACIA) |
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No separation step Enzyme activity deminishes when Antigen-Antibody binding occurs |
Homogenous Assay |
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Requires a separation step |
Heterogenous assay |
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Medium to which an antigen or Antibody may be attached |
Solid-Phase |
GCPM 1. Glass Beads 2. Cellulose Membranes 3. Polyesterene Tubes 4. Microtiter Plates |
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Enzyme Immunoassay: Enzyme Labels |
1. Glucose Oxidase 2. G6PD 3. Horseradish Peroxidase 4. Alkaline phosphatase 5. B-galactosidase |
GGHAB
Horseradish Peroxidase - most common |
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In Enzyme Immunoassay change in absorbance is measured using |
Spectrophotometry |
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Enzyme Immunoassay: Types |
1. Competitive 2. Non-competitive/Indirect 3. Capture Assay/Sandwich 4. Enzyme Multiplied Immunoassay Technique (EMIT) |
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Enzyme labeled antigen competes with unlabeled patient antigen for a limited number of Antibody-Binding sites
Enzymatic activity is inversely proportional to analyte concentration |
Competitive Enzyme Immunoassay |
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Enzyme-labeled secondary antibody does not participate in the initial Antigen-Antibody binding Enzyme activity is directly proportional to the analyte concentration |
Non-competitive/Indirect Enzyme Immunoassay |
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Antigen captured must have multiple epitopes
Prone to hook effect Enzyme activity is directly proportional to Antigen Concentration |
Capture Assay/Sandwich |
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Unexpected fall in amount of measured analyte when extremely high concentration is present |
Hook Effect |
In case of prozone, majority of binding sites are filled, and the remainder of the patient antigen has no place to bind |
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A Homogenous Assay
Antigen is labeled with an enzyme tag |
Enzyme Multiplied Immunoassay Technique (EMIT) |
When Antibody binds to specific determinant sites on the antigen, the active site on the enzyme is blocked, resulting to loss of Enzyme activity |
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Enzyme labels with the highest turnover and high sensitivity |
1. Alkaline Phosphatase 2. Horseradish Peroxidase |
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Labels used in Fluorescent Immunoassay |
1. Fluorescein Isothiocyanate (FITC) 2. Tetramethylrhodamine Isothiocyanate (TRITC) 3. Phycoerythrin 4. Europium (B-naphthyl fluoroacetone) 5. Lucifer Yellow VS |
Extremely specific and sensitive |
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Label that has green fluorescence against a red background |
Fluorescein Isothiocyanate (FITC) |
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Label that has a Red fluorescence |
Tetramethylrhodamine Isothiocyanate (TRITC) |
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Fluorescence can be measured using |
1. Fluorometer 2. Fluorescent Microscope (Requires UV light) 3. Flow Cytometer 4. Spectrofluorometer |
FLUO |
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Atoms emit light of _______ wavelength and ________ energy during transition to the ground state |
Longer, Lower |
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Disadvantages of Fluorescent Immunoassay |
1. Autoflurescence (False Inc) 2. Quenching (Falsely Dec) |
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Fluorescent Immunoassay : Types |
1. Direct Immunofluorescent Assay 2. Indirect Immunofluorescent Assay 3. Inhibition Immunofluorescent Assay 4. Fluorescence Polarization Immunoassay |
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Unknown Antigen is Fixed to microscope slide Fluorescent labeled Antibody is added directly to unknown antigen |
Direct Immunofluorescent Assay |
Slide read using Fluorescent Microscope |
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Blocking test in which antigen is first exposed to unlabeled Antibody, then to labeled Antibody, and is finally washed then examined |
Inhibition Immunofluorescent Assay |
No Fluorescence If the unlabeled and labeled antibodies are both homologous to the antigen |
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Based on the change in polarization of Fluorescent tag emitted from a labeled molecule when it is bound by Antibody
Degree of Fluorescence Polarization is inversely proportional to analyte Concentration |
Fluorescence Polarization Immunoassay (FPIA) |
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First type of Immunoassay developed |
Radioimmunoassay |
Label: Radioisotpes 125I -most popular (60 days Half-life) 131I 3H |
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Radioactivity is measured by |
Scintillation Counter |
Crystal scintillation counter: Gamma
Liquid scintillation counter: Beta |
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Disadvantages of Radioimmunoassay |
1. Health Hazard 2. Disposable problems 3. Short shelf life 4. Need for expensive equipment |
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Measures total IgE |
Radioimmunosorbent Test (RIST) |
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Measures Allergen specific IgE |
Radioallergosorbent Test (RAST) |
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Radiolabled antigen competes with unlabeled patient antigen for limited number of Antibody-Binding sites |
Competitive Binding Assays |
Radioactivity is inversely proportional to Antigen Concentration |
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Analyte measured is sandwiched between 2 antibodies |
Non-competitive Immunoradiometric Assays (IRMA) |
Radioactivity is directly proportional to the analyte concentration |
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dsDNA is heated to 95C to separate the DNA into single strands |
DENATURATION |
PCR |
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DNA is cooled to 52C to allow primers to bind to complimentary sequences on separate DNA strands |
ANNEALING |
Bind/Anneal |
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At 72C, the heat-stable DNA polymerase (Taq polymerase) binds to 3' end of Each primer and Synthesizes a new strand of DNA |
ELONGATION or Extension |
Polymerization |
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Taq polymerase is a thermostable polymerase isolated from the bacterium |
Thermus aquaticus |
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Marker for Hepa B Active Infection
First marker to appear
Important Marker in screening blood donors |
HBsAg (surface antigen) Australia Antigen |
Sero Markers for HepB: 1. HBsAg (Surface) - First Marker to appear - ACTIVE INFECTION 2. HBeAg (Envelope) - High deg of Infectivity, High Vertical transmission risk - Active Replication 3. HBcAg (Core) - Detected thru liver biopsy - Undetectable in serum (Envelope) 4. IgM anti-HBc - Current or recent infection - CORE WINDOW 5. IgG anti-HBc - LIFELONG MARKER 6. Anti-HBe - RECOVERY - Convalescence Marker (Good Prognosis) 7. Anti-HBs - IMMUNITY |
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Hepa B Marker that denotes High degree of infectivity & high vertical transmission risk Active replication of the virus |
HBeAg (envelope antigen) |
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Lifelong Marker for Hepa B |
IgG anti-Hbc |
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Hepa B Marker detected only through biopsy of the infected liver Undetectable in Serum because of the viral envelope that masks it |
HBcAg (core) |
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Hepa B Marker that denotes current or recurrent infection Useful in detecting infection during the core window period |
IgM anti-HBc |
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Hepa B Marker that denotes Immunity to hepatitis B |
Anti-HBs |
Protective titer of >10 mIU/mL of Serum |
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Hepa B Marker that denotes Recovery from infection
Convalescence maker (good prognosis) |
Anti-HBe |
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Hepa B Marker that denotes acute, atypical or occult hepatitis Viral load test to monitor effectiveness of therapy |
HBV DNA |
Highly correlated w/: 1. Whole virus replication 2. Potential Infectivity of the patient |
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Hepa A marker that shed in the feces during incubation period and early acute infection |
HAV Antigens |
Sero Markers for HAV: 1. HAV Antigens - Shed in Feces during incubation period and early acute inf 2. IgG anti-HAV - Immunity 3. Total anti-HAV - Immunity 4. HAV RNA - Detection in clinical, food, water samples |
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Hepa A marker for Immunity to hepatitis A |
1. IgG anti-HAV 2. Total anti-HAV
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Hepa A marker for detection of HAV in clinical, food, water samples |
HAV RNA |
Nucleic Acid Testing |
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Hepa C marker for current or past Hepa C infection |
Anti-HCV |
Sero Markers for HCV: 1. anti-HCV - CURRENT OR PAST INF 2. HCV RNA - Viral Load test to monitor effectiveness of therapy - Determine Genotype |
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Hepa C marker for Viral Load Test to monitor effectiveness of therapy |
HCV RNA |
Determine genotype |
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Hepa D marker for acute or chronic infection |
IgM anti-HDV |
Sero Markers for HDV: 1. IgM anti-HDV - ACUTE OR CHRONIC INF 2. IgG anti-HDV - RECOVERY 3. HDV RNA - ACUTE - Viral Load Test |
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Hepa D marker for recovery from infection |
IgG anti-HDV |
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Hepa D marker for active infection Viral load test to monitor effectiveness of therapy |
HDV RNA |
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Hepa E Markers for current infection |
1. IgM anti-HEV 2. IgG anti-HEV 3. HEV RNA |
IgG anti-HEV - also a marker for past infection |
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Non-Chronic Hepatitis |
Hepa A and E |
Chronic: B, C, D |
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From Family Picornaviridae (RNA)
Transmission: Fecal Oral
Incubation: 28 days
May be transmitted by clotting factor concentrates |
Hepa A |
HEPATITIS: 1. HEPA A - Picornaviridae (RNA) - 28 DAYS 2. HEPA B - Hepadnaviridae (DNA) - 60-90 Days 3. HEPA C - Flaviviridae (RNA) - 7-8 Wks 4. HEPA D - Delta Virus (RNA) 5. HEPA E - Herpesviridae or Caliciviridae (RNA) - 3-8 Wks |
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From Family Hepadnaviridae (DNA) Transmisaion: Parenteral, Sexual, Perinatal Incubation: 60-90 days |
Hepa B |
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From Family Flaviviridae (RNA)
Transmission: Parenteral, Sexual, Perinatal
Incubation: 7-8 wks
POST TRANSFUSION HEPA |
Hepa C |
Surrogate tests: 1. Inc ALT 2. (+) Anti-HCV (SPECIFIC) |
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From Genus Delta Virus (RNA)
Transmission: Mostly Paranteral, also sexual, perinatal
HBV Inf Required |
Hepa D |
Co-infection: Simultaneously Superinfection: Sequentially |
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From Family Herpesviridae or Caliciviridae
Transmission: Fecal, Oral
Incubation: 3-8 wks Fulminant liver failure in pregnant women |
Hepa E |
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Complete HBV that causes infection |
Dane particle |
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First Generation test for HBsAg |
Ouchterlony Double Diffusion |
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2nd Generation test for HBsAg |
1. Counterelectrophoresis 2. Complement Fixation 3. Rheophoresis |
CCR |
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3rd Generation tests for HBsAg |
1. Radioimmunoassay 2. Reverse Passive hemagglutination 3. Reverse Passive Latex Agglutination 4. Sandwich/Capture EIA |
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A Retrovirus containing RNA and reverse transcriptase Targets Helper CD4+ T Cells |
Human Immunodeficiency Virus |
HIV-1 formerly called: 1. Human T-Cell Lymphotrophic Virus type III (HTLV-III) 2. Lymphadenopathy-associated virus (LAV) 3. AIDS-Associated Retrovirus (ARV) |
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HIV 1 Strains Coreceptors
Along with CD4 to bind & enter target cells
1. X4 2. R5 3. R5X4 |
1. X4 (T-tropic) - CXCR4 - Expressed on T cells
2. R5 (M-Tropic) - CCR5 - Expressed on Monocytes, T cells
3. HIV-1 R5X4 - Both CCR5 and CXCR4 |
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Causative agent of AIDS in the United States and Europe |
HIV-1 |
HIV-2 - endemic in West Africa, less pathogenic, lower rate of transmission |
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3 major routes for transmission of HIV: |
1. Intimate Sexual Contact 2. Parenteral (Infected Blood, Body Fluids) 3. Perinatal (Infected mother to infant) |
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Cornerstone of screening procedures for HIV |
ELISA |
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Standard treatment for HIV |
HAART (High active antiretroviral therapy) |
Regimen involving combination of atleast 2 drug classes |
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HIV protein product Responsible for binding to CD4+ Receptor T Cell |
gp120 |
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First to appear during infection with HIV |
Antibody to p24 |
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Positive Result for Western Blot is the presence of 2 out of 3 major bands: |
1. p24 2. gp41 3. gp120/160 |
gp160 - a precursor protein that is cleaved from gp120 and gp41 |
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HIV Ab identification screening tests |
1. ELISA 2. Agglutination Tests 3. Dot-Blot Testing |
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HIV Ab Identification Confirmatory tests |
1. Western Blot/Immunoblot 2. Immunofluorescence |
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Gold Standard for CD4+ T Cell Enumeration |
Immunophenotyping w/ Flow Cytometry |
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HIV-AIDS CD4+ T cell Count |
<200 cells/mm3 |
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Used to identify HIV infection during the window period before Antibody is detectable |
p24 antigen detection |
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HIV Test used to determine viral load and development of drug-resistant strains |
HIV Nucleic Acid Test |
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Quantitative Test for HIV Nucleic Acid Most sensitive test for diagnosis before seroconversion |
VIRAL LOAD TESTS |
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Preffered method for HIV detection in infants and children younger than 18 months |
PCR |
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Solid-Phase Indirect-Assay using viral lysate antigens from HIV-1
Detects antibodies to HIV-1 only |
First Generation ELISA |
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Indirect binding Assay using highly purified recombinant or synthetic antigens from both HIV-1 & HIV-2 Detects antibodies to HIV-1 & HIV-2 |
Second Generation ELISA |
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Sandwich technique Detects HIV Antibodies of different Ig Class, including IgM |
Third Generation ELISA |
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Detects: 1. HIV-1 Antibodies 2. HIV-2 Antibodies 3. p24 Antigens |
Fourth Generation ELISA |
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According to CDC, when ELISA yields a positive result it should be |
1. Retested in duplicate by the same ELISA test 2. 2 out of 3 specimens are reactive, the results must be confirmed by a more specific test |
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False Positive ELISA Test |
1. Heat inactivation of Serum prior to testing 2. Repeated freezing/thawing 3. Autoreactive antibodies 4. Multiple pregnancies 5. Severe Hepatic Dse 6. Passive Ig Administration 7. Recent exposure to certain vaccines 8. Malignancies |
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False Negative ELISA test |
1. Collection of the test Serum prior to seroconversion 2. Immunosuppressive Therapy or replacement transfusion 3. Hypogammaglobulinemia 4. Technical Errors 5. Patient Harbors a genetically diverse, recombinant strain of HIV |
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Product of gag (group antigen gene) |
p55 |
p55 - a precursor protein which forms core Proteins: 1. p6 2. p9 - core binding protein 3. p17 - inner surface of envelope 4. p24 - Core coat for nucleic acids |
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Products of pol (polymerase) gene |
1. Reverse Transcriptase (p66, p51) - Transcribes RNA to DNA
2. Integrase (p31) - Inserts Viral DNA to Host DNA
3. Protease (p10) - Cleaves protein precursors
4. RNAse |
RIPR |
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Products of env (envelope) gene |
1. gp160 - Cleaved to form gp120 & gp41
2. gp120 - Binds to CD4+ Tcells
3. gp41 - Transmembrane protein associated with gp120 |
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Product of tat gene that activates transcription of HIV provirus |
p14 |
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Product of vif gene which is an infectivity factor |
p23 |
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Product of nef gene that enhances HIV replication |
p27 |
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Product of vpr gene functions for integration of HIV DNA into host genome |
p15 |
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Product of vpu gene functions for Viral assembly and budding |
p16 |
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Product of rev gene that transports viral mRNA to the cytoplasm of host cell |
p19 |
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Best indicator of the immediate state of immunologic competence of a patient with HIV infection |
CD4+ T Cell Count |
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Stage 3 HIV disease |
One or more specific opportunistic illness has been diagnosed |
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AIDS defining Illnesses |
1. CMV Retinitis 2. HIV Encephalopathy 3. Invasive Cervical Cancer 4. P. jirovecii Pneumonia 5. Burkitt's lymphoma 6. Esophageal Candidiasis 7. Kaposi's Sarcoma |
CHIPBEK |
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Self-Limiting disease caused by EBV |
Infectious Mononucleosis |
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EBV Transmission: |
1. Saliva 2. Blood Transfusion - IM Post Perfusion Syndrome 3. Transplacental |
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Cells infected by EBV |
B Cells |
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Cell seen in EBV
Enlarged Lymphocyte with atypical nuclei BALLERINA SKIRT |
Downey Cell |
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Other diseases associated w/ EBV |
1. Burkitt's Lymphoma 2. Nasopharyngeal Carcinoma |
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Antibodies that will cross react with any member of a group of similar antigens that can be found in unrelated animals or microorganisms |
Heterophile Antibody |
Heterophile Antigens - So similar that an Antibody built to one will cross react with the others |
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HETEROPHILE ANTIBODY Characteristics |
1. Reacts w/ horse, ox, sheep RBC'S 2. Adsorbed by beef erythrocytes not adsorbed by Guinea Pig Kidney Cells 3. Does not react w/ EBV-specific antigens |
IgM that usually appears during acute Phase of IM |
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Hemagglutination test designed to detect Heterophile antibodies in patient Serum when mixed with antigen-binding sheep rbc's Useful screening but cannot determine Specificity |
PAUL-BUNNEL Screening Test |
Serological Tests for IM: 1. Paul-Bunnel Screening Test 2. Davidson Diff Test 3. Monospot Test/ Rapid Diff Test |
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Normal Titer for PAUL-BUNNEL Screening Test |
</equal to 56 |
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Heterophile Antibody titer is reported as the |
Reciprocal of the highest dilution showing Agglutination |
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Distinguishes between Heterophile antibodies associated with infectious Mononucleosis, Serum Sickness, or Frossman Antigen |
Davidson Differential Test |
Absorption w/ 1. Guinea Pig 2. Beef RBC
Agglutination 1. Sheep RBC |
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Not absorbed: Guinea Pig Kidney Cells Absorbed: Beef RBC |
IM |
Absorbed: (-) Agglutination w/ Sheep RBC Not Absorbed: (+) Agglutination w/ Sheep RBC |
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Absorbed: 1. Guinea Pig Kidney Cells 2. Beef RBC |
Serum Sickness |
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Absorbed: Guinea Pig Kidney Cells Not absorbed: Beef RBC |
Forssman |
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Horse RBC's are agglutinated by Heterophile antibodies of IM |
Monospot Test/ Rapid Differential Slide Test |
Also a differential but require absorption of the patient's Serum |
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EBV marker detectable early in the course of infection |
IgM anti-VCA (Viral Capsid Antigen) |
EBV Sero Markers: 1. IgM anti-VCA - Early in the course of Inf 2. IgG anti-VCA - After onset of signs & symptoms - Can be present for life 3. IgG anti-EAD - Active Inf 4. IgG anti-EAR - Found in serum of very young children but not in young adults during acute stage 5. EBNA - Found in nucleus of ALL EBV-infected cells - Does not stimulatr ab until after incubation period of IM |
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EBV marker found in the nucleus of All EBV-infected cells |
EBNA (Epstein Barr Nuclear Antigen) |
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EBV marker found the Serum of very young children but not in young adults during acute stage |
IgG anti-EA-R (Early Antigen-Restricted) |
EA-R |
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EBV marker detected after onset of signs and symptoms Can be present for life |
IgG anti-VCA |
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EBV marker which strongly indicates active infection
Not consistent Indicator of the disease |
IgG anti-EA-D |
EA-D (Early Antigen-Diffuse) |
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Syphilis Transmission |
1. Sexual Contact 2. Direct Blood Transmission 3. Transplacentally |
Syphilis previously called Great Pox or Evil Pox |
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Drug of Choice for Syphilis |
Penicillin G |
Salvarsan (606)/Arsphenamine Used as treatment for Syphilis in the 1900s |
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Specific anti-treponemal antibodies in early or early untreated latent syphilis are predominantly |
IgM antibodies |
Early immune response rapidly followed by appearance of IgG antibodies, soon become predominant |
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Greatest elevation of IgG concentration is seen in __________ stage of Syphilis |
Secondary |
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Non-specific antibodies against the protein antigen group common to pathogenic sphirochetes |
NONTREPONEMAL Antibodies |
Often called REAGIN Antibodies
Produced by infected patients against components of their own or other mammalian cells |
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Other Infectious Diseases with Reagin |
1. Measles 2. Chicken Pox 3. Hepatitis 4. IM 5. Leprosy 6. TB 7. Leptospirosis 8. Malaria 9. Rickettsial Dse 10. Trypanosomiasis 11. Lymphogranuloma venereum |
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Non-infectious Diseases with Reagin |
1. Autoimmune Disorders 2. Drug Addiction 3. Old Age 4. Pregnancy 5. Recent Immunization |
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Stages of Syphilis: |
1. PRIMARY (EARLY) STAGE - Hard Chancre 2. SECONDARY STAGE - Condylomata Lata (Generalized Rash) 3. LATE LATENT STAGE - No Signs & Symptoms - Sero Tests Pos 4. TERTIARY STAGE - Gummas - Nuerosyphilis |
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Parenchymatous Neurosyphilis |
1. |
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Direct detection of Spirochetes |
1. Dark Field Microscopy 2. Fluorescent Antibody Test |
Dark Field Microscopy: Observe for corkskrew motility of spirochetes |
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Tests which detects reagin
Non-specific only for screening |
NONTREPONEMAL Tests |
Reagin/Nontreponemal/Anticardiolipin/Antilipoidal Ab Nontreponemal Serological Tests: 1. VDRL 2. RPR |
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Process in which reagin reacts with lipid antigens from animal tissue (beef heart) |
FLOCCULATION |
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Biologic False Positive in Nontreponemal Tests |
1. SLE 2. RA 3. IM |
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Tests that detects Antibodies to T. pallidum |
Treponemal Tests |
Treponemal Serological Test: 1. T. pallidum Immobilization (TPI) Test 2. Fluorescent Treponemal Ab Absorption Test 3. Agglutination Tests - Hemagglutination Treponemal Tests for Syph (HATTS) - T. pallidun Hemagglutination Assay (TPHA) - Microhemagglutination Assay For Ab to T. pallidum (MHA-TP) - T. pallidum Particle Agglutination (TPPA) |
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NEW WORLD to OLD WORLD |
SYPHILIS |
SMALL POX: OLD WORLD to NEW WORLD |
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1st diagnostic test developed in 1906 which uses Cardiolipin as Antigen |
WASSERMAN TEST |
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A qualitative/quantitative slide Flocculation test |
Vinereal Disease Research Laboratory (VDRL) Test |
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Specimen for VDRL Test |
50 uL serum/csf Heated for 30 mins at 56C |
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Antigen composition in VDRL Test |
1. 0.3% Cardiolipin 2. 0.9% Cholesterol 3. 0.21% Lecithin
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Uses slides with ceramic rings 1. Cardiolipin - main reacting component 2. Cholesterol - enhances reacting surface of cardiolipin 3. Lecithin - removes anti-complement activity of cardiolipin |
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Antigen delivery needles (Hamilton Syringe) |
1. Qualitative Serum VDRL 18g - 60 drops Ag suspension/mL 2. Quantitative Serum VDRL 19g - 75 drops Ag suspension/mL 23g - 100 drps Ag suspension/mL 3. CSF VDRL 21/22g - 100 drops/mL |
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VDRL: Rotator: Serum |
4 mins, 180 rpm |
VDRL: Rotator: CSF 8 mins, 180 rpm |
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A positive VDRL Test on spinal fluid is diagnostic of |
Neurosyphilis |
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A modified VDRL Test with Charcoal Particles |
Rapid Plasma Reagin (RPR) Test |
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Antigen used in RPR is similar to VDRL but with addition of |
1.Charcoal - for macroscopic visibility reaction 2. Thimerosal - Preservative 3. EDTA 4. Choline Chloride - inactivate complement |
"ChaThiECho" Antigen Delivery Needle: 20g, 60 drops Ag suspension/mL |
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Specimen for RPR |
50 uL serum No Heating |
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RPR: ROTATOR |
8 mins, 100 rpm |
Same reporting w/ VDRL |
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Treponemal Antibody, in the presence of complement, inhibits normal movement of actively motile treponemes extracted from Testicular chancre of rabbit |
Treponema Pallidum Immobilization (TPI) Test |
Treponemal Test reference |
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Patient Serum is heat inactivated and made with a sorbent consisting of nonpathogenic treponemes (Reiter strain) which is used to remove cross reactivity with Treponemes other than T. pallidum |
Fluorescent Treponemal Antibody Absorption Test |
Nichol's strain of T. pallidum have been fixed to slides used for the test Ab from pt + Nichol's strain |
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Agglutination Test for Syphilis which uses glutaraldehyde-stabilized Turkey RBC's |
Hemagglutination Treponemal Test for Syphilis (HATTS) |
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Agglutination Test for Syphilis which uses Tanned Sheep RBC's coated with antigen from Nichol's Strain |
T. pallidum Hemagglutination Assay (TPHA) |
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Agglutination Test for Syphilis which uses formalinized, Tanned sheep RBC's sensitized with antigen from Nichol's Strain |
Microhemagglutination Assay for Antibodies to T. pallidum (MHA-TP) |
Similar to TPHA but performed in Microtiter Plates |
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Agglutination Test for Syphilis which uses gelatin Particles instead of RBC's sensitized with T. pallidum antigens |
T. pallidum Particle Agglutination (TPPA) |
TPPAG |
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Other Treponema-Associated Diseases in Humans 1. Yaws 2. Pinta 3. Bejel 4. Rabbit Syphilis |
1. Yaws: T. pertenue 2. Pinta: T. carateum 3. Bejel: T. pallidum endemicum 4. Rabbit Syphilis: T. cuniculi (WASSERMAN Nonreactive) |
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Complement Inactivation: |
1. Heating to 56C for 30 mins 2. If >4hrs have elapsed after inactivation, reinactivate by heating to 56C for 10mins |
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Non-treponemal Tests |
1. VDRL slide test 2. RPR 3. Plasmacrit 4. (TRUST) Toluidine Red Unheated Serum Test 5. (USR) Unheated Serum Reagin 6. (RST) Reagin Screen Test |
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Treponemal tests |
1. TPI 2. T. pallidum complement fixation 3. Reiter Protein complement fixation 4. Fluorescent Treponemal Antibody Test 5. Fluorescent Treponemal Antibody Absorption Test 6. T. pallidum Hemagglutination 7. Direct Fluorescent Antibody Test |
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A Febrile Antibody Test used for detection of antibodies in 1. Typhoid Fever 2. Brucellosis 3. Tularemia |
Widal Test |
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Clinically Significant Titer in Widal Test |
>/= to 160 |
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A Febrile Antibody Test that uses cross-reacting P. vulgaris & P. mirabilis antigens to diagnose Rickettsial Infection |
Weil-Felix Test |
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Antigens from P. vulgaris |
OX-2 OX-19 |
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Antigen from P. mirabilis |
OX-K |
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Disease Caused by: 1. R. prowazekki 2. R. typhi 3. R. rickettsia 4. R. akari 5. R. tsutsugamushi 6. B. quintana 7. C. burnetti |
1. R. prowazekki - Epidemic Typhus - Brill-Zinsser Dse 2. R. typhi - Endemic Typhus - Murine Typhus 3. R. rickettsia - Rocky Mountain Spotted Fever 4. R. akari - Rickettsialpox 5. R. tsutsugamushi - Scrub Typhus 6. B. quintana - Trench Fever 7. C. burnetti - Q Fever |
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Malarial Test which detects Parasitic Lactic Dehydrogenase which is produced by viable Parasites |
OptiMAL |
Different isoforms of lactate Dehydrogenase are present in different spp. |
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pLDH can be detected when thare are |
100-200 parasites/uL of blood |
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Malarial Test that detects P. falciparum Histidine Rich protein 2 antigen |
MalaQuick Standby Malaria Test |
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Gold Standard for detecting Rickettsial Antibodies |
1. Indirect Fluorescence Assay (IFA) 2. Microimmunofluorescence Assay (MICRO-IF) |
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Test for Group A Streptococcal Infection which is based on Neutralization of hemolytic activity of Streptolysin O |
Antistreptolysin O Titer |
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Titer for ASO is reported as the |
Reciprocal of the highest dilution showing no hemolysis |
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ASO Titer is expressed using |
1. Todd Units 2. International Units |
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Normal Titer in ASO |
</equal to 166 Todd Units |
Moderately Elevated in Adult: >/equal to 320 Todd Units Moderately Elevated in Children:
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Group A Streptococcal Infection Test which is based on Neutralization of depolymerizing activity of DNAse B |
Anti-DNAse B Test |
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1 |
1 |
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2 |
2 |
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3 |
3 |
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Slide Coagulation screening Test for detection of antibodies to several Streptococcal antigens |
Streptozyme Test |
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