Serology

Study of bodily fluids

 Standard Analysis of Blood
 Questions about crime scene blood:
 Is the substance blood?
 Is the blood human?
 Can the blood be associated with a particular
individual?
 Is the substance blood?
 Kastle-Meyer Color test
 Phenolphthalein, hydrogen peroxide, and
suspected blood stain are mixed; positive
reaction indicated by a deep pink color
 Note: This test is a presumptive or preliminary test
for blood; meaning that it is not definite. Other
testing must be done to determine if the substance
is truly blood.
 Note: Substances other than blood may produce
the pink color change—this is called a false
positive result.
 Is the substance blood? Con’t
 Other tests for blood include the Hemastix
and benzidine color test.
 Hemastix—has been found to be very useful
by field investigators—a dipstick moistened
with distilled water is placed in contact with
the suspect blood—a green color change
indicates a positive result
 Benzidine has been phased out, because it is
a known carcinogen.
 Is the substance blood? Con’t
 Luminal Test
 A presumptive test for blood that produces light when
it reacts with blood rather than color.
 Large areas can be quickly screened for blood, by
spraying luminal reagent on a suspect item or area.
 Sprayed objects must be in a darkened area while
being viewed for the emission of light (luminescence)
 Luminal test is extremely sensitive—it is capable of
detecting blood stains that have been diluted up to
300,000 times.
 Dose not interfere with subsequent DNA testing
 Is the blood human?
 Precipitin Test
 This test is based on the theory of antigen-antibody
reactions.
 If you inject an animal with human blood, the animal
will create antibodies that can then be recovered and
used to test for various human antigens including
human blood antigens
 The recovered antibodies are called human antiserum
 To test for human blood, human antiserum and an
unknown blood are put together—if the blood is
human a precipitate will form.
 Can the blood be associated with a
particular individual?
 A-B-O types, enzymes, and other factors
 Until the mid 1990’s, characterization of bloodstains
included determining A-B-O types, enzyme markers
such as PGM (phosphoglucomutase), and other
factors
 These tests add statistical information with which to
reduce the number of possible sources of the
bloodstain
 Because antigens, and enzymes occur independently
of one another, the probability of a dried blood stain
having a particular combination of these factors is
determined by the product of their distribution in the
population
 Can the blood be associated with a
particular individual? Con’t
 Statistical Example:
 Blood stain found to be of type A—
corresponds to 42% percent of population
 Stain contains PGM-1—corresponds to 58% of
population
 The combined effect of just these two factors,
using the product rule is 42% x 58% or 24 %
 Clearly the more factors, the smaller the frequency
of occurrence in the population
 Can the blood be associated with a
particular individual? Con’t
 DNA Analysis
 Since the 1990’s DNA analysis has replaced
the forensics reliance on the routine
characterization of bloodstains to a particular
individual using blood factors.
 DNA analysis gives very accurate
individualization results.
 Stain Patterns of Blood
 The distribution and appearance of
bloodstains and spatters can be used to
interpret and reconstruct the events that
must have occurred to produce the
bleeding.
 Stain Pattern Analysis
General Conclusions Con’t

 Surface texture: The harder and less

porous the surface, the less spatter
results.
 Direction of Travel: The direction of travel
of blood can be discerned by the stain’s
shape—the pointed end always faces its
direction of travel
 Stain Pattern Analysis General
Conclusions Con’t
 Impact Angle: A drop of blood striking a surface
at a right angle gives rise to a nearly circular
stain; as the angle decreases, the stain becomes
more elongated in shape
 Origin of Blood Spatter: Origin in a two
dimensional configuration can be determined by
establishing lines through the long axis of
several individual bood stains. The intersection
of the lines represents the point from which the
blood emanated.
Example of Origin of Blood Spatter
 Nature of Blood
 Blood refers to complex mixture of cells,
enzymes, proteins, and inorganic
substances
 Liquid portion is called plasma/serum
composed of mainly water, accounts for 55
% of the blood content
 Solid portion is made up of RBCs, WBCs
and platelets
 Nature of Blood Con’t
 Antigens
 Reside on the RBC’s surface and impart blood-
type characteristics to the cell
 Stimulate the body to produce antibodies
against it
 More than 15 blood antigen systems have
been identified, including A-B-O and Rh
systems
 Note: These systems are used forensically to
identify an individual
 Nature of Blood Con’t
 Antibodies
 Destroy or inactivate a specific antigen
 Found in the blood serum
 For every antigen there is a corresponding
antibody
 Antibodies only react with a specific antigen
 The reaction causes agglutination (clumping)
 ABO System
Blood Type Antigens/RBC Antibodies/Serum

A A Anti-B

B B Anti-A

AB AB Neither anti-A or B

O Neither A nor B Both anti-A and B

 Blood Typing
Anti-A Anti-B Antigen Blood Type
serum + Serum + Present
blood blood
+ _

_ +

+ +

_ _
 Principles of Heredity
 Transmission of hereditary material is
accomplished by means of microscopic
units called genes
 The gene is the basic unit of heredity. It is
positioned on chromosomes. All human
cells contain 46 chromosomes, mated in
23 pairs. (Only exception: reproductive
cells)
 Principles of Heredity Con’t
 During fertilization, a sperm and egg
combine so each contributes
chromosomes to form the new cell
(zygote)
 Hence, the new individual inherits genetic
material from both parents.
 Just as chromosomes come in pairs, so do
the genes they bear.
 Principles of Heredity Con’t
 Genes that govern a given characteristic
are similarly positioned on the
chromosomes inherited from the mother
and father.
 Alternate forms of genes that influence a
given characteristic (ex. Eye color) are
aligned with one another on a
chromosome pair are called alleles
 Principles of Heredity Con’t
 Inheritance Rules
A gene pair of two similar genes—is
homozygous—no problem determining a
person’s outward characteristics
 A gene pair of two different genes—is
heterozygous—one gene is dominant, the
other recessive—person’s outward
characteristics depend on the dominant gene
 Punnet Square
Father’s Genotype
O O

Mother’s
A AO AO
Genotype

B BO BO

Note: genotype is the pair of allele genes

Phenotype is the outward physical
expression of the genes
 Analysis of Punnet Square
 50 % of offspring are genotype AO
 50 % of offspring are genotype BO
 Because O is recessive 50% of offspring
are type A blood and 50 % are type B
blood
NOTE: From this example, we can see that
no blood group gene can appear in a child
unless it is present in at least one of the
parents.
 Paternity Testing
 Many cases of disputed paternity can be
resolved when suspected parents and
offspring are related according to their
blood group systems
 Paternity Example

Father’s Genotype
O O

Mother’s
A AO AO
Genotype

B BO BO

Ifthe offspring had been found to be blood type AB

would the suspect father be cleared?
 Paternity Example
Father’s Genotype
O O

Mother’s
A AO AO
Genotype

B BO BO

What if the child had been found to be blood type A or

type B? What could we say about the suspect father?
 Paternity Testing Con’t
 Note: Routine paternity testing involves
characterizing many blood factors other
than A-B-O. Using these other factors the
probability of establishing paternity
becomes better than 90. Once DNA
analysis is done the odds become better
than 99%.
 Forensic Characterization of Semen
 A large number of cases involve sexual offenses,
making it necessary to examine exhibits for the
presence of seminal stains.
 The forensic examination of semen is a two-step
process:
 First, the semen must be located
 Second, tests must be performed to prove its
identity—it may even be tested for blood type
 Note: Semen is checked for blood type because many
individuals are secretors
 Secretors
 80% of individuals are classified as secretors
 This means that their blood-type antigens are
also found in high concentrations in most body
fluids
 Ex- Saliva, semen, vaginal secretions, and
gastric juices
 **Saliva & semen have higher concentrations of
A & B antigens than blood!!
 Semen Location and Identification

 There are two methods of locating semen:

 First,semen is often readily visible on a fabric
because it has a stiff crusty appearance—at
best this method is unreliable though
 Second, the acid phosphate color test can
locate and at the same time identify semen.
 Acid Phosphate Color Test
 Acid phosphate is an enzyme secreted by
the prostate gland into seminal fluid that
is in far greater concentrations than those
found in any other body fluids.
 Its presence can be detected two ways:
 1. Contact with a special acidic solution and
fast blue B dye shows a color change
 2. Contact with a solution of MUP fluoresces
under UV light.
 Acid Phosphate Test, Con’t
 Large areas or fabrics can be quickly searched
by moistening filter paper with the detector
solutions and rubbing over the suspect area
 Some substances will give a false-positive result
(fruit juices, contraceptive creams, and vaginal
secretions), but none of these substances will
react with the speed of seminal fluid.
 Note: A reaction of less than 30 seconds is a strong
indication of semen.
 Microscopic Examination of Semen

 The presence of spermatozoa (telltale

shape, head-tail, etc.) absolutely indicates
the presence of semen.
 Rapid stirring of a fabric-stain-water
mixture will transfer a small amount of
sperm into the water. A drop is then
taken, dried, stained and looked at under
the microscope.
 Microscopic Problems
 Locating sperm with the microscope is not that
easy for three reasons:
 1. Sperm are very brittle when dried and easily
disintegrate. They are also hard to remove from cloth
material.
 2. Sexual crimes may involve males who have an
abnormally low sperm count, condition called
oligospermia.
 3. Or males who have no sprem in their seminal fluid,
condition called aspermia—often due to increasing
numbers of vasectomies.
 Detecting Semen, Con’t
 A positive result for acid phosphate and a
negative result for semen give ambiguous
results—In this case, p-30 or prostate
specific antigen (PSA) is used to detect
seminal plasma.
p30 prostate specific antigen test
 Formation of a visible line between the
wells of anti p-30 and semen extract–
proves the stain is seminal
 Collection of Rape Evidence
 Outer and undergarments are placed in separate
paper bags, while victim stands on a piece of
paper. The paper is then folded and collected.
 Bedding or the object upon which assault took
place should be submitted to the lab.
 Rape vicitim must be subjected to a medical
examintion Collected items include pubic hairs,
vaginal swabs, blood, fingernail scrapings,
clothing and urine)
 Collection Con’t
 Suspect must also undergo an exam.
Routine items collected include clothing,
hairs, penile swab and blood samples
 Timeline
 Living or motile sperm may survive 4 to
six hours in the vagina of a living person.
 Nonmotile sperm may be found in a living
female for up to three days and
occasionally up to six days
 Intact sperm (sperm with tails) are not
normally found 16 hours but have been
found as late as 72 hours after
intercourse.
 Important Considerations Based
Upon Timeline
 How long ago was the person raped?
 When did voluntary sexual activity last
occur prior to the sexual assault?
 Bloodis taken from any consensual partner
having sex with the victim within 72 hours of
the assault.