Forensic Biology I: Quiz 2/Exam 2 Study Guide

The scientific name for the Takayama crystals are called___________ ______________

__________________ __________________

pyridine ferroprotoporphyrin.

The four ingredients of Takayama reagent:

1. Water

2. Pyridine

3. Saturated glucose

4. 10% NaOH

The two characteristics of the Takayama crystals.

1. Pink

2. Feather-shaped

Sensitivity of the Takayama crystal test.

It is not very sensitive, but it is good up to 1:20 dilution.

Five different tests for species identification:

1. Identification of red blood cell morphology

2. Precipitin ring test (not currently used)

3. Immunodiffusion with specific antigens and antibodies:

A. Single Immunodiffusion

B. Double Immunodiffusion

4. Immunoelectrophoresis

5. ABA card hematrace

A species test identifies...

the species of the blood sample.

The measurement of RBCs as precisely as possible in smears made from fresh blood as well as in reconstituted stains is called __________________

micrometry.

RBCs of mammals are ______________________ and non mammals are ________________.

enucleated; nucleated.

Human and Duck blood can be differentiated by the presence or absence of_______________ in the Red blood corpuscles.

a nucleus

T/F: Micrometry is a process by which the white blood cell sizes and shapes are measured as precisely as possible in smears made from fresh blood as well as in reconstituted stains for species identification.

False

Antiserum

A serum that contains an antibody and reacts against antigen.

T/F: Antibodies are proteins.

True

The precipitin ring test is based on the reaction between __________ and ___________.

antigen; antibody.

These two people described the double immunodiffusion technique:

1. Elek (England)

2. Orjan Ouchterlony (Sweden).

In single immunodiffusion, the material that diffuses is __________________.

antigen (goat serum)

In double immunodiffusion the materials that diffuse are _______________ and__________________.

the antigen (stain extract); antibody (antiserum).

The material that is impregnated in the agarose medium in single immunodiffusion is _____________.

goat antiserum.

The formation of precipitin band formation in precipitin ring test indicates a _______________ reaction (positive, negative, false positive, false negative).

positive

Orjan Ouchterlony's name is widely associated with __________________________.

Ouchterlony double immunodiffusion.

Ouchterlony described the agar gel immunodiffusion in this book called_________________.

Handbook of Immunodiffusion and Immunoelectrophoresis.

In the Ouchterlony double immunodiffusion test, it is called 'double' diffusion because______________.

both the antigen and antibody (antiserum) diffuse.

Three types of immunodiffusion patterns (identity, non-identity, and partial identity):

1. Identical = peak

2. Partially identical = Spur

3. Non-identical = X

In the Ouchterolony plate, the identity of two proteins are indicated by_________________.

fusion band.

T/F: In the Ouchterlony double immunodiffusion test, the non-identical proteins are indicated by a spur formation.

False

In the Ouchterlony double immunodiffusion test, the spur formation indicates __________ identity.

partial

T/F: In the Ouchterlony double immunodiffusion test, it is possible to see two or more band formations between the antigen well and the antibody well.

True

The two other animals/species (besides human) that react with the ABAcard hematrace test device are __________________ and ______________________.

primates; ferrets

The higher concentration of hemoglobin in a sample causes a weak reaction in the ABAcard hematrace test device and this effect is called ___________________________________________.

high dose hook effect.

The precipitin ring test, immunodiffusion test and the ABA card test devices are based on the reactions of _______________ and __________________.

antigen; antibody.

The name of the antigen detected in the ABA card one step kit for the detection of human blood is _________________.

human hemoglobin.

The principle involved in the ABA card one step test kit:

Based on the antigen-antibody reaction.

Human hemoglobin from the sample will react with the anti-human hemoglobin antibody present in the test and form a colored band in the test area.

There are ____________ (#) different types of antibodies present in the ABA card test kit.

three

The names of antibodies present in the ABA card one step test kit for human blood:

1. Mobile Anti-Human Hemoglobin Antibody

2. Immobilized Anti-Human Hemoglobin Antibody

3. Immobilized Immunoglobulin Antibody

What is high does hook effect? What happens during this process? How do we correct this problem in the species identification test?

1. The high-dose hook effect is caused by a high concentration of human hemoglobin.

2. During this effect, free hemoglobin migrates to the test area and blocks the binding of the antigen-antibody complex to the immobilized antibody.

3. You can correct this effect by diluting the sample.

The company that manufactures the Aba card one step test kit is ______________.

Abacus Diagnostics

Antibody raised against human hemoglobin will react with __________________.

human hemoglobin.

T/F: The antibodies raised against deer heart muscle will react with human heart muscle.

False

The negative test result in ABA card is indicated by the absence of colored band in this area. (control area; test area; sample area; test and control area)

Test area

The absence of colored band in the control area in the aba card indicates: a: human blood; b: non human blood: c: invalid test result.

Invalid test result.

The reaction of Ferret blood with the Aba card test device is considered ___________________ (false positive or false negative results).

false positive.

The reaction of ferret blood with aba card test device is due to:

ferret has some common amino acid sequence homology with human hemoglobin.

T/F: The aba card is sensitive enough to give a positive reaction with cattle blood.

False

The antibodies present in the control region of the Aba card hematrace test kit is____________.

immobilized immunologlobin antibody.

T/F: Each red blood cell contains two antigens.

False

Antigens are proteins, that stimulate the body to produce _____________ against it.

antibodies

There are ____________ (#) reactive sites in each antibody.

two

There are ___________ (#) heavy chains and ___________ (#) light chains present in an antibody.

two; two

The heavy chains and light chains are held together by _____________ bonds.

disulphide

T/F: The amino acid sequence of the variable portion of an antibody will differ among different antibodies.

True

Structure of an antibody and the parts.

T/F: Immunoelectrophoresis involves the movement of antigens and antibodies in an electrical field.

True

T/F: Different proteins (antigens) can be separated and identified in immunoelectrophoresis.

True

The pattern that will be seen on the Ouchterlony test plate with the following antigen-antibody configuration:

antigen: a - antigen: a

antibody- a

Peak shaped identical.

The pattern that will be seen on the Ouchterlony test plate with the following antigen-antibody configuration:

antigen: a - antigen: b

antibodies- a and b

X-shaped not identical.

ABAcard Hematrace cards: Sample

Human Hemoglobin (Antigen)

ABAcard Hematrace cards: Sample Area (Sample)

Mobile Anti-Human Hemoglobin Antibody

ABAcard Hematrace cards: Test Area (Test)

Immobilized Anti-Human Hemoglobin Antibody

ABAcard Hematrace cards: Control Area (Control)

Immobilized Immunoglobulin Antibody

ABAcard Hematrace cards: Positive Result

1. Human hemoglobin (antigen) was added to the test device.

2. Mobile antihuman hemoglobin antibody conjugated to a pink dye in the card.

3. Human hemoglobin binds with mobile antihuman hemoglobin antibody.

4. The mobile antigen-antibody complex is formed.

5. The mobile antigen-antibody complex migrates to the test area.

6. Immobilized antihuman hemoglobin antibody - in the test area.

7. The mobile antigen-antibody complex binds with the immobilized antihuman hemoglobin antibody forming an antibody-antigen-antibody complex.

8. Visualized by the formation of a pink color band.

9. The mobile antihuman hemoglobin antibody not connected to the human hemoglobin (antigen) migrates to the control area.

10. Immobilized immunoglobulin antibody - in the control area.

11. The mobile antihuman hemoglobin antibody binds to the immobilized immunoglobulin antibody forming an antibody-antibody complex.

ABAcard Hematrace cards: Negative Result

The mobile antigen-antibody complex migrates to the test area but doesn't bind to immobilized antihuman hemoglobin antibody.

Instead, it migrates to the control area and binds to the immobilized immunoglobulin antibody forming an antibody-antibody complex.

ABAcard Hematrace cards: Invalid Result

The mobile antigen-antibody complex doesn't migrate to the test area.