Species Identification (Exam 2) Flashcards
What 5 tests can be used for species identification?
1) ID of RBC morphology
2) Precipitin ring test
3) Immuno-diffusion w/ specific antigens & antibodies
4) Immunoelectrophoresis
5) ABA card hematrace
Define micrometry.
Measurement of RBCs as precisely as possible in smears made from fresh blood as well as reconstituted stains
What does a species specificity test determine?
If a blood sample is human or animal
What is the difference between mammal and nonmammal RBCs?
Mammalian RBCs are not nucleated
In which 4 nonmammal groups will you find nucleated RBCs?
1) Birds
2) Reptiles
3) Amphibians
4) Fish
Aside from meaning “red blood cell,” RBC can also mean…
Red blood corpuscle
The precipitin ring test relies on the formation of a band between these 2 proteins.
Antigens & antibodies
What are the 2 types of immunodiffusion?
Single & double
Which 2 people described the double immunodiffusion process (HINT: the former is English and the latter is Swedish)?
Elek & Orjan Ouchterlony
In single immunodiffusion, the material that diffuses is the…
Antigen
In double immunodiffusion, the two materials that diffuse are the…
Antigen & antibody
In the beginning of single immunodiffusion, Bechold (1905) allowed { } to be diffused into a gelatin medium.
Goat anti/serum
How do you make a 1% agarose solution?
1g agarose : 100mL buffer
What does the presence of a band indicate in the precipitin ring test in terms of results?
Positive
Who began using agar gel plates for double immunodiffusion?
Orjan Ouchterlony
What was the name for Ouchterlony’s book describing the process of agar gel immunodiffusion?
Handbook of Immunodiffusion & Immunoelectrophoresis
What does the term “double” refer to in Ouchterlony double immunodiffusion?
The stain extract (antigen) and the antiserum placed in adjacent agarose wells
How is the identity of two proteins indicated by an Ouchterlony plate?
A precipitin fusion band between the 2 proteins
What does spur formation indicate in Ouchterlony double immunodiffusion?
Non- or partial identity
Who makes the ABAcard Hematrace cards?
Abacus Diagnostics
Aside from humans, what two species react with the hematrace test device?
Ferrets & primates
What is the cause of the high dose hook effect?
High concentrations of human hb
What is the effect of the high dose hook effect?
Weakened reaction in the hematrace test and, therefore, no pink color formation
How much of a sample do you need for the hematrace cards?
Just a trace amount
What are the 3 types of species specificity tests that are based on antigen-antibody reactions?
1) Precipitin ring
2) Double immunodiffusion
3) ABA hematrace
What is the mechanism of action behind the high dose hook effect?
Free hb migrates to the test area and blocks the binding of Ab/Ag complex to the immobilized antibody
What are the 3 components of a hematrace card?
1) Positive control
2) Negative control (TEST)
3) Sample
What kind of antibody will you find in the positive control of an ABA card?
Immobilized immunoglobin
What kind of antibody will you find in the negative control (TEST portion) of an ABA card?
Immobilized human hemoglobin
What kind of antibody will you find in the sample portion of an ABA card?
Mobile human hemoglobin
Two bands present on an ABA card indicate…
Positive result
One band present, which is in the positive control, of an ABA card indicates…
Negative result
No band in either the positive or negative control of an ABA card indicates…
Invalid result
Why does ferret hemoglobin show up as a false positive on an ABA card?
Ferret hb contains an amino acid sequence that is homologous to the sequence found in human hb
