Forensic Serology

and DNA Analysis

◼ SFS3. Obtain, evaluate, and communicate
information relating to biological evidence in
forensic investigations.
c. Construct an explanation to distinguish the difference
between human and animal blood.
d. Plan and carry out an investigation to analyze the
physics of bloodstain patterns.
e. Plan and carry out an investigation involving DNA
processing and analysis.
What is forensic serology?
◼ Forensic serology is the detection,
classification and study of various
bodily fluids such as blood, semen,
fecal matter and perspiration, and
their relationship to a crime scene.
◼ A forensic serologist may also be
involved in DNA analysis and
bloodstain pattern analysis.
The Nature of Blood
◼ Blood is a specialized bodily fluid that
is composed of two substances –
cells and plasma
◼ It is responsible for delivering
nutrients and oxygen to and
transporting wastes and carbon
dioxide from cells, tissues, and
organs.
The Nature of Blood
◼ The average human body has about
5 liters of blood (less for females,
more for males).
◼ Blood accounts for 8% of your weight
◼ Oxygenated blood is bright red, while
deoxygenated blood is darker red due
to the nature of hemoglobin
The Nature of Blood
◼ Erythrocytes (red blood cells)
Contain hemoglobin and distribute
oxygen
Account for ~93% of all blood cells
◼ ~5 billion cells/L
Represent ~25% of all cells in the body
Over 2 million made every second
Live for about 120 days before recycled
Hematocrit is a measure of RBC volume
(45% of whole blood)
The Nature of Blood
◼ Leukocytes (white blood cells)
Function in the immune system
Five types categorized by immune
target and function
Account for ~0.1% of all blood cells
◼ ~7 billion cells/L
Livefor 3-4 days before recycled
Main component of pus
The Nature of Blood
◼ Thrombocytes (platelets)
Responsible for blood clotting
Produced by fragmentation of larger
stem cells
Account for ~7% of all blood cells
◼ ~250 billion cells/L
“Live”for 5-9 days before recycled
Form long fibers to trap RBCs and form
scab
 THROMBOCYTE
(platelet)

ERYTHROCYTE
(red blood cell)

LEUKOCYTE
(white blood cell)
The Nature of Blood
◼ Plasma is the fluid portion of blood
and accounts for 55% of the blood
content.
◼ It is mostly water, and it contains the
clotting factors, other proteins,
hormones, and various chemicals.
◼ Serum is the liquid that separates
from the blood when a clot is formed
(serum = plasma – clotting factors)
The Nature of Blood:
Basic Immunology
◼ An antigen is any substance that
causes your immune system to
produce antibodies against it.
◼ The substance may be from the
environment or formed within the
body.
◼ The immune system will kill or
neutralize any antigen that is
recognized as a foreign and
potentially harmful invader.
Blood typing is…
◼ the classification of blood based on
the presence or absence of inherited
antigenic substances on the surface
of RBCs
◼ More than 30 blood antigen systems
have been identified (over 600
individual antigens!), but the ABO and
Rh systems are the most important.
 ABO system was discovered by Landsteiner in
1901 (Nobel Prize in 1930)
Blood Typing
◼ Type A RBCs have A antigens, and
the blood plasma has anti-B antibodies
◼ Type B RBCs have B antigens, and
the blood plasma has anti-A antibodies
◼ Type AB RBCs have both A and B
antigens, and the blood plasma has no
antibodies
◼ Type O RBCs have no antigens, but
the blood plasma has both anti-A and
anti-B antibodies
Blood Typing
◼ Rh factor is determined primarily by
the presence of the D antigen.
◼ RBCs with the D antigen are Rh +
◼ RBCs without the D antigen are Rh -
Blood Typing
◼ Because you have
pairs of
chromosomes,
your genotype is
composed of two
alleles for the ABO
blood group gene.
◼ Your phenotype
results from your
specific genotype.
How common are the blood types?
In the US
In general…
•African Americans and
Hispanics are more likely
to have Type O blood.
•Caucasians are more
likely to have Type A
•Asians are more likely to
have Type B and AB
blood
Blood Typing
◼ When an antibody
comes into
contact with its
target antigen,
clumping, called
agglutination,
occurs.
Blood Donation and Transfusion
◼ Because of the immune response in
blood, donation and transfusion is
dependent on donor and recipient
blood type.
◼ Recipient blood should have NO
antibodies that could react to the
donor antigens, otherwise the blood
would agglutinate in the recipient’s
vessels, leading to rejection and
death.
Blood Donation and Transfusion
◼ So, which blood type is BEST for
donation?
◼ Which type recipient would have the
easiest transfusions?
Immunoassay Techniques
◼ An immunoassay is a biochemical
test that measures the presence or
concentration of a substance in
solutions that frequently contain a
complex mixture of substances.
◼ A number of immunological assay
techniques are commercially
available for detecting drugs through
antigen-antibody reaction.
Example of Immunoassay
Bloodstains at the Crime Scene
When blood is found there are four
questions to ask:

1. Is it really blood?
2. Is the blood human?
3. From whom did the blood come?
4. How did the blood get there?

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Is it really blood?
Kastle-Meyer Color Test
◼ Stain is mixed with phenolphthalein reagent
and then hydrogen peroxide.
◼ If it is blood, the sample will turn a bright
pink color due to hemoglobin’s oxidative
effects.
◼ Can be a false positive since some
substances have the same oxidative effects,
so a confirmatory test would be needed.
25
Is it really blood?
Hemastix Strips
◼ Strip is moistened with distilled water and
placed on blood stain.
◼ If blood is present, the strip will turn green.

26
Is it really blood?
Luminol Test
◼ Luminol reagent is sprayed onto a surface,
in as dark of a room as possible.
◼ If blood is present a luminescent glow will
show in the area of the blood, as it reacts
with the iron in the hemoglobin.
◼ The Luminol test is very sensitive and can
detect bloodstains diluted up to 10,000
times!
◼ Can be false positive with feces, urine, or
27
some bleaches
28
Is the blood human?
◼ To determine whether a blood sample is
from a human or animal source, samples
are tested with anti-human serum.
 The forensic test consists of collecting the
blood sample in a test tube containing an
animal-derived serum containing antibodies
against human blood.
 If an insoluble complex of precipitin (this
would be visible as agglutination) occurs, the
test is positive for human blood.
From whom did the blood come?
◼ Bloodtyping and/or DNA testing
(discussed later) can determine this.
How did the blood get there?
◼ Dr.Herbert MacDonell, founder of the
Bloodstain Evidence Institute in
Corning, New York, is acknowledged
as the foremost pioneer in analyzing
blood patterns and has been
internationally acclaimed for over five
decades for his forensic expertise.
MacDonell’s Observations for Blood:

Surface texture is of paramount

importance in the interpretation of
bloodstain patterns, and correlations
between standards and unknowns
are valid only if identical surfaces
are used.
In general, the harder and less
porous the surface, the less spatter
results.
33
MacDonell’s Observations for Blood:

The direction of travel of blood

striking an object may be discerned
by the stain’s shape.
The pointed end of a bloodstain
always faces its direction of travel.
35
MacDonell’s Observations for Blood:

It is possible to determine the impact

angle of blood on a flat surface by
measuring the degree of circular
distortion of the stain.
A drop of blood striking a surface at
right angles gives rise to a nearly
circular stain; as the angle
decreases, the stain becomes
elongated in shape.
37
MacDonell’s Observations for Blood:

The origin of a blood spatter in a two-

dimensional configuration can be
established by drawing straight lines
through the long axis of several
individual bloodstains.
The intersection or point of
convergence of the lines represents
the point from which the blood
emanated.
39
Other issues…
◼ Wetblood has more value than dried
blood because more tests can be run.
For example, alcohol and drug content
can be determined from wet blood only.
◼ Bloodbegins to dry after 3-5 minutes
of exposure to air.
As it dries, it changes color towards
brown and black.
 saturation
Blood Spatter Types
◼ Passive
Created by the force of gravity

drops drip flow pool

Blood Spatter Types
◼ Projected
Occurs when some form of energy is
transferred to the blood source

Low velocity Medium velocity High velocity

impact impact impact
Blood Spatter Types
◼ Projected

cast-off arterial spurting

Blood Spatter Types
◼ Projected

back spatter expiratory

Blood Spatter Types
◼ Transfer
Produced when an object with blood
comes in contact with an object or
surface that does not have blood
contact wipe swipe
DNA Collection & Analysis
◼ Investigators gather samples from the
crime scene and from suspects and
then analyze it for a set of specific
DNA regions or markers.
◼ A match of one marker is not usually
unique, but if a sample matches four
or five markers, there is a very good
chance it is a match.
DNA Collection
◼ DNA is collected at crime scenes in a
variety of ways:
 Sterile swabs (for moist substance)
 Scalpels (for scraping dried substance)
 Collection kits (samples from suspects
or living victims)
DNA Collection
◼ DNA samples can be from:
 Saliva
 Blood
 Hair
 Skin
 Finger or toe nails
 Tooth with root material
DNA Extraction
DNA Analysis
◼ Several DNA technologies are used in
DNA analysis:
RFLP
PCR
STR
mtDNA Analysis
RFLP: Restriction Fragment Length
Polymorphism
◼ Analyzes variable lengths of DNA
fragments
◼ Used in the development of DNA
fingerprinting (Dr. Alec Jeffreys, 1985)
◼ Not used as much anymore because…
 itrequires a large DNA sample
 the processing is slow and tedious
 samples degraded by the environment do not
work well with RFLP
The basic RFLP technique:
◼ A sample of DNA is combined with
restriction enzymes that break apart the
strands at specific locations
◼ The resulting fragments are sorted
according to their lengths by gel
electrophoresis

Gel Electrophoresis Simulation

PCR: Polymerase Chain Reaction

◼ Used to make millions of exact copies of

DNA from a biological sample
◼ Allows very small samples to be analyzed,
such as a sample of a few skin cells
◼ Must be very careful about contamination
in this process
The basic PCR technique:

◼ DNA strand is heated; two pieces separate

◼ Each half is used as a template to build a
new complementary strand

PCR Simulation
55
STR: Short Tandem Repeat
◼ Evaluates specific regions (loci) within
nuclear DNA
◼ FBI uses 13 standard specific STR regions
for CODIS
◼ The purpose of establishing a core set of
STR loci is to ensure that all forensic
laboratories can establish uniform DNA
databases and, more importantly, share
valuable forensic information.
Mitochondrial DNA Analysis

◼ Used for samples that cannot be analyzed

using RFLP or STR
◼ Uses DNA extracted from mitochondrion
rather than nuclear DNA
◼ Especially useful in old cases and old
samples
DNA Collection & Comparison
◼ What happens after the samples are collected?
A DNA profile is created….how??
◼ Markers are found by designing small pieces of DNA
(probes) that will seek out and bind to complementary DNA
sequences. This creates a distinct pattern. Again, one
marker is not usually unique, but with four or five regions the
match is likely
 The DNA profiles are compared with samples from
suspects to find possible matches.
 If there are no suspects, a national database called
CODIS may be used to find potential suspects.
DNA Collection & Comparison

◼ More on CODIS:
 Stands for Combined DNA Index System
 National network that helps identify leads for
crimes with no suspects
 Three tiers: Local (LDIS), State (SDIS),
National (NDIS)
 Uses 13 DNA regions that vary from person to
person
 Looks for matches at more than one location
on a genome for more accurate results
Sources of DNA at Crime Scenes

◼ Cool table at:

 http://www.dna.gov/basics/evidence_collectio
n/identifying
Sources of DNA at Crime Scenes

◼ Examples of sources from real cases:

 Saliva on the stamp of a stalker’s threatening letter
 Skin cells shed on a ligature of a strangled victim
 Perspiration on a baseball cap discarded by a rapist
was compared with the saliva swabbed from a bite
mark on a different rape victim
 A single hair (without the root) found deep in a
victim’s throat
 Maggot digestive contents
How can DNA evidence be
planted?
◼ Sneezing or coughing over evidence
◼ Person touches their mouth, nose or other
part of the face and then touches the area
that may contain the DNA to be tested.
◼ Scene personnel can deposit hairs, fibers,
or trace material from their clothing
◼ Wind can carry in contaminants
Other Uses of DNA

◼ Paternity Testing and Proving Family

Relations
◼ Identification of John or Jane Does
◼ Study of evolution and ancestry
◼ Studying Inherited Disorders