BASIC SERO TECHNIQUES, PRECIP'N, AGGLU'N, LABELED IA'S, PCR

• If testing cannot be performed immediately < 72 hrs - serum is stored at _

  2-8° C
• If testing cannot be performed immediately > 72 hrs - serum is stored at _

  -20° C
• Inactivation of serum – heating of serum at _ for _ _

  56° C, 30 mins
• Reinactivation of serum (when 4 hrs has lapsed) – heating of serum at _ for __

  56° C, 10 mins
• indication of Relative Concentration, and maybe expressed as fraction or a ratio

  DILUTION
• represents the ratio of concentrated or stock material to the total final volume of a solution

  DILUTION
• 3 TYPES OF DILUTION

  Simple, Compound, Serial Dilution
• Serial dilution is also called

  Two-Fold dilution
• Defined as the reciprocal of the highest dilution of the patient’s serum in which the antibody is still detectable or had produced a positive result.

  Antibody Titer
• Antibody Titer is determined by performing _ _ and adding equal amount of antigen to each dilution

  Serial Dilution
• Ab + soluble Ag = insoluble complexes


  Precipitation
• Agglutination

  Ab + particulate Ag = cellular aggregations
• sum total of all the forces that exist between multivalent Ab and multivalent Ag

  AVIDITY
• initial force of attraction that exist between a single Fab and epitope of an antigen

  AFFINITY
• where optimum precipitation/agglutination occurs

  ZONE OF EQUIVALENCE
• Prozone phenomenon = _ _

  excess in Abs
• Postzone phenomenon = _ _

  excess in Ags
• Widely used to quantify immunoglobulins

  Nephelometry
• Nephelometry measures the _ _ _ _ in a solution containing antibody–antigen complexes

  amount of light scattered
• Turbidimetry measures the _ _ _ _ in a solution of antibody–antigen complexes

  decrease in light intensity
• [TURBIDIMETRY] the _ the light intensity = the _ the concentration of complexes

  lower, higher
• Precipitation by Passive Immunodiffusion Support media:

  gel, agar, agarose
• Uses a tube or column with intervening column for precipitation reaction

  Oakley-Fulthorpe
• Single Diffusion in One Dimension aka

  Oudin
• Double Diffusion in One Dimension aka


  Oakley-Fulthorpe
• What is the difference between Oudin and Oakley-Fulthorpe?

  Intervening column
• In Radial Immunodiffusion, the Ab is incorporated with the _ _ while the Ag is added to _ - _ _


  agar gel, pre cut walls
• the antigen of Mancini aka _ _ is allowed or not allowed to diffuse completely?

  Endpoint method, allowed
• The antigen of Kinetic method aka _ & _ is allowed or not allowed to diffuse completely?

  not allowed
• Fahey & McKelvey formula

  d = LOG of Ag concentration
• Mancini formula

  d2 = Ag concentration
• What is the incubation period of IgG & IgM?

  24 hours, 50-72 hours
• What is the incubation period of Fahey & McKelvey?

  18 hours
• Double Diffusion in Two Dimension (+) Result

  Distinct pattern of precipitation
• Single Diffusion in Two Dimension (+) Result

  Precipitin ring
• Double Diffusion in Two Dimension aka

  Ouchterlony Double Diffusion
• [Patterns of ppt'n] Spur formation

  Partial identity
• [Patterns of ppt'n] Non-identity

  Crossed-line
• [Patterns of ppt'n] Smooth curve

  Serologic Identity
• Rocket Immunoelectrophoresis


  RID + electrophoresis