1. Identification Using Blood Groups & Stains
Forensic Serology
Identification Using Blood Groups & Stains
This presentation contains graphic pictures.

2. Forensic Serology
Serology – term used to describe a broad range of laboratory tests using reactions of blood serum and body fluid
The serology section of a forensic laboratory may deal with any or all of the following:
blood typing
characterization of unknown blood
stain patterns for crime reconstruction
paternity testing
semen identification in rape cases
DNA techniques used for identification

3. The Composition of Blood
Blood is a mixture of many components:
cells inorganic substances (salts)
enzymes water
proteins
55 % of blood content is plasma – mostly water and substances dissolved in it
Most of the solid materials (by weight) are cells
red blood cells, RBCs (erythrocytes)
white blood cells (leukocytes)

4. Blood Typing Example
A sample of unknown blood is mixed with three anti-sera samples:
Tube 1 (Anti-A): No reaction
Tube 2 (Anti B): No reaction
Tube 3 (Anti Rh): Cloudy reaction
In terms of the A-B-O and Rh systems, what type blood is the sample?
The ABO antigens are not confined to RBC; 80% of Americans are secretors, found on other cells

5. Blood Typing Example
A sample of unknown blood is mixed with three anti-sera samples:
Tube 1 (Anti-A): No reaction
Tube 2 (Anti B): No reaction
Tube 3 (Anti Rh): Cloudy reaction
In terms of the A-B-O and Rh systems, what type blood is the sample?
The ABO antigens are not confined to RBC; 80% of Americans are secretors, same antigens found on other cells
O positive

6. Forensic Characterization of Bloodstains
Three questions that must be answered by the forensic investigator:
1) Is it blood?
Benzidine Test - Benzidine is carcinogen so not used much Benzidine + Blood Stain + Hydrogen peroxide = pink color
Kastle-Meyer Test
Hemastix (o-tolidine, TMB) – detects peroxide activity of hemoglobin
Luminol Test
Microcrystaline test
2) Is it human blood?
Precipitin Test
3) Can it be associated with an individual?

7. H2O2 is a by-product of cellular reactions… (remember that?)

8. The HEME in your RBC, acts as the peroxidase...

9. It breaks up the H2O2...

10. The O combine with the KM, our phenolphthalein color-indicator.

11. Luminol - the most sensitive chemical test that is capable of detecting bloodstains diluted to as little as 1 in 100,000
Red blood cells contain hemoglobin (Hb) – the protein responsible for transporting oxygen
Each Hb contains four iron (Fe) containing hemes
Heme structure

12. Microcrystalline tests
Microcrystalline tests, such as the Takayama and Teichmann tests
depend on the addition of specific chemicals to the blood so that characteristic crystals will be formed.

13. Precipitin Test

14. The cell membrane of RBCs has molecules in its surface that act as identification badges so immune system can recognize it as a normal component.
Foreign
Normal
IMMUNE SYSTEM
Red Blood Cell
CHEMICAL MARKERS
ANTIBODIES

15. Blood Antigens
On the surface of red blood cells are chemical structures (proteins) called antigens
The presence of antigens allow a living system to recognize foreign biological substances
Antigens also impart specific characteristics to the red blood cells. These characteristics can be partly used to identify individuals
Human red blood cells have more than 15 different antigen types that have been discovered

16. Blood Typing
Blood typing involves determination of the antigens present on an individual’s RBCs
The two most common blood typing systems used are the A-B-O method and the Rh method
type A blood – contain “A” antigen on RBCs
type B blood – contain “B” antigen on RBCs
type AB blood – contains both A and B antigens
type O blood – contain no A or B antigens
Rh+ blood – contain Rh antigen
Rh- blood – no Rh antigen

17. Blood Typing
Blood also contains antibodies – proteins that recognize and bind to certain, specific antigens
Blood Type Antigens on RBC Antibodies
A A anti-B
B B anti-A
AB AB none
O none both A & B

18. Blood Typing
When blood containing a specific antigen is mixed with blood containing the corresponding antibodies, the blood clots
The clots can then be removed by the host system

19. Blood Typing
Human serum containing specific antibodies can be purchased
Separately mix a drop of unknown blood sample with drop of each anti-serum
Reaction between blood and anti-serum
No reaction between blood and anti-serum

20. Opener:
Watch the video clip and describe how the analyst is able to determine if blood is present.
Watch this video clip and explain what data can be obtained by looking at bloodstain patterns

21. Blood Splatter Analysis
Categories of Bloodstains:
Passive (dripping)
Transfer (smearing)
Projected
Projected Bloodstains occur in shootings, trauma from blunt weapons, hacking, or slashing attacks.

22. Projected Bloodstain Analysis
Two Important Determinations:
a. direction of splatter
b. angle of impact with surface
sin  = (width drop / length drop)

23. Forensic Characterization of Semen
Many crimes involve sexual misconduct
Normal males, upon ejaculation, release seminal fluid. This fluid is a mixture of components:
water
spermatozoa
enzymes
inorganic salts
Semen at a crime scene (clothing, bed sheets, carpet, cushions) can be located and identified by three common methods:
microscope examination to find sperm
acid phosphatase test
p30 (also called prostrate specific antigen) immunoassay test

24. Forensic Characterization of Saliva
Saliva is a mixture of many components:
99% water
Mucin (protein helps in swallowing)
Amylase (enzyme to help digest carbohydrates)
Cheek cells (good for DNA)
Adults produce liters of saliva/day and it is not uncommon at crime scenes (especially involving bite marks).
A simple test for saliva involves mixing starch, iodine, and a sample of the presumed saliva together. Starch and iodine are a deep blue color when mixed together. The amylase breaks down starch, however, and the color fades (takes about oC).