Forensic Serology

DR. dr.Ahmad
Dr.Ahmad
Yudianto,SpF.,MKes.,SH
Yudianto,dr.SpF.,SH.,M.Kes

Department of Forensic Medicine & Medicolegal

Forensic Science
of Programme
Study
Faculty of Medicine,
Airlangga
University
School of Postgraduad,
Airlangga University
2010
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

BLOOD TYPING

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

Luminol

From What Species does the Blood

Morphological differences
Originate?
Precipitin Test Precipitin Test
Blood sample & Human anti serum
Agglutinate
Capillary tube
Gel diffusion
Electrophoretic method

Precipitin Test

Blood analysis

Conventional Serological Analysis

Analysis of Proteins, Enzymes & Antigens
Blood Typing
Traditional A-B-O & Rh Factor typing using
known anti known anti-serum or known cells
serum or known cells
Immunoassay 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 individuals 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

anti-B

anti-A

AB

AB

none

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 antisera 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 antisera 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? O positive
The ABO antigens are not confined to RBC; 80% of
Americans are secretors, same antigens found on
other cells

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)

BLOODSPATTER

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).