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

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

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)

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

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

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?

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

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

It breaks up the H2O2...

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

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

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.

Precipitin Test

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

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

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

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

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

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

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

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

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

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

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 1.0-1.5 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 15 mins @ 37 oC).