Forensic Serology
DR. dr.Ahmad Yudianto,SpF.,MKes.,SHDepartment of Forensic Medicine & Medicolegal
Faculty of Medicine, Airlangga University2010
Dr.Ahmad Yudianto,dr.SpF.,SH.,M.Kes
Forensic Science of Programme StudySchool of Postgraduad, Airlangga University
2013
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
Detection and Identification of Bloodstain
• Histological, Histochemical & Cytological• Chemical test : - Preliminary test (Colour test): - Benzidine test - Guaiac Test - Phenolphthalin test - Conclusive test (Crystal test): - Teichmann test - Takayama test - Physico-Chemical test : - Spectroscopy - Polargraphy - Elctrophoresis - Chromatography• Immunologcal Methods--- Precipitine test• Serological Identification• Electrophoretic Techique• Autoanalyser Technique
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)
Composition of Blood
• Elements (46%) – Cells
• Erythrocytes • Leukocytes
– Polymorponuclear » Neutrophils » Eosinophils » Basophils
– Mononuclear » Monocytes » Lymphocytes
– Platelets • Plasma (54%)
– Water, Electrolytes, Nutrients – Proteins
• Albumin • Immunoglobulins (antibodies) • Fibrinogen
Forensic Characterization of Bloodstains
Three questions that must be answered by the forensic investigator:
1) Is it blood?
Benzidine Test
Kastle-Meyer Test
Hemastix (o-tolidine)
Luminol Test
2) Is it human blood?
Precipitin Test
3) Can it be associated with an individual?
Detection - Presumptive Tests
• Erythrocytes contain Haemoglobin (Hb)
• Haemoglobin has Peroxidase activity & is used
in tests to identify the presence of blood.
– Benzidine Colour Test ( 1: 200,000)
– Phenolphthalein ( Kastle-Meyer Colour Test)
• Hemastix
• Luminol Test
– Sensitivity:1:5,000,000
• Microcrystalline Tests
Luminol
• Red blood cells contain hemoglobin (Hb) – the protein responsible for transporting oxygen
• Each Hb contains four iron (Fe) containing hemes
Heme structure
How does Luminol detect blood?
• Blood is slightly alkali
• Hemoglobin is a ferrous compound
• Metals are a catalyst in the reaction between an alkali and Luminol
Hemoglobin
From What Species does the Blood Originate?
• Morphological differences
• Precipitin Test Precipitin Test
Blood sample & Human anti serum
Agglutinate
– Capillary tube
– Gel diffusion
– Electrophoretic method
Blood analysis
Conventional Serological AnalysisConventional Serological Analysis
• Analysis of Proteins, Enzymes & Antigens
Blood TypingBlood Typing
Traditional A-B-O & Rh Factor typing using known anti known anti-serum or known cells serum or known cells
Immunoassay TechniquesImmunoassay Techniques
- Presence of particular drug
- Sub-typing iso-enzymes of polymorphic enzymes in erythrocytes or serum
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 methode and the Rh methode
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
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
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 SemenMany 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 SalivaSaliva 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).