Chapter 8 Serology

1
Chapter 8Serology
Out damned spot! Out, I say Heres the smell
of the blood still, All the perfumed of Arabia
will not Sweeten this little hand. Oh, Oh,
Oh! William Shakespeares Lady Macbeth, in
Macbeth
Correlation coefficient
http//noppa5.pc.helsinki.fi/koe/corr/cor7.html
2
Learning Objectives
Students should be able to

• List the A-B-O antigens and antibodies found in
  the blood for each of the four blood types A, B,
  AB and O.
• Understand and describe how whole blood is typed.
• List and describe forensic tests used to
  characterize a stain as blood.

3
Learning Objectives
Students should be able to

• List and describe forensic tests used to
  characterize a stain as blood.
• Understand the concept of antigen-antibody
  interactions and how it is applied to species
  identification and drug identification.

4
Learning Objectives
Students should be able to

• Contrast chromosomes and genes.
• Use the Punnett square to determine the genotypes
  and phenotypes of offspring.

Fathers genotype
O O
A
Mothers genotype
B
5
Learning Objectives
Students should be able to

• List the laboratory tests necessary to
  characterize seminal stains.
• Explain how suspect blood and semen stains are
  properly preserved for laboratory examination.

UV light makes seminal fluids glow brightly
6
Learning Objectives
Students should be able to

• Describe the proper collection of physical
  evidence in a rape investigation.

7
Learning Objectives
Students should be able to

• List the A-B-O antigens and antibodies found in
  the blood for each of the four blood types A, B,
  AB and O.
• Understand and describe how whole blood is typed.
• List and describe forensic tests used to
  characterize a stain as blood.

8
Forensic Serology

• Forensic serology is the study of blood, saliva,
  semen or sweat in manners pertaining to the law.
• From 1950 to the early 1990s, forensic serology
  was a most important part of lab procedures.
• With the development of DNA techniques, more
  time, money, and significance was placed in
  developing DNA labs.
• However, with limited funds and the time required
  for DNA testing, many labs still use many of the
  basic serology testing procedures.

9
Historical Perspective of Blood Typing

• In 1901, Karl Landsteiner discovered that there
  are four different types of human blood based on
  the presence or absence of specific antigens
  found on the surface of the red blood cells.
• By 1937, Landsteiner and Weiner reported the
  discovery of the Rh factor by studying the blood
  of the Rhesus monkey. 85 of Caucasians, 94 of
  Black Americans and 99 of all Asians are Rh
  positive.
• More than 100 different blood factors have
  been identified.

10
I. The Nature of Blood
11
Blood Characteristics

• Plasma is the fluid portion of the blood (55)
• Cells (45)
• Erythrocytes are red blood cells. They are
  responsible for oxygen distribution.
• Leukocytes are the white blood cells they are
  responsible for cleaning the system of foreign
  invaders.
• Thrombocytes or platelets are responsible for
  blood clotting
• Serum is the liquid that separates from the blood
  when a clot is formed.
• Antigens, usually proteins, are located on the
  surface of RBCs and are responsible for
  blood-type characteristics.

12
Human Blood

• Red blood cells are most numerous 5 to 6 million
  per mm3
• White blood cells are larger and less numerous
  5000 to 10,000 per mm3
• Platelets are tiny, cellular fragments 350,000
  to 500,00 per mm3

13
Blood Terminology

• ABO blood groupsbased on having an A, B, both or
  no antigens on red blood cells
• Rh factormay be present on red blood cells
  positive if present and negative if not
• Antigena substance that can stimulate the body
  to make antibodies. Certain antigens (usually
  proteins) located on the red blood cells
  membrane account for blood type.
• Antibodya substance that reacts with an antigen
• Agglutinationclumping of red blood cells will
  result if blood types with different antigens are
  mixed

14
Blood Typing

• More than 15 blood antigen systems have been
  identified, but the 2 most important are
• the A-B-O system
• A, B, AB and O blood types
• the Rh system
• Rh positive and Rh negative

15
Blood Typing

• The identity of each of the four A-B-O blood
  groups can be established by testing the blood
  with anti-A and anti-B sera.
• For every antigen there is a specific antibody
  that will react with it to form clumps known as
  agglutination.
• If serum containing anti-B is added to RBCs
  carrying the B antigen, they will immediately
  react (clump together).

16
Blood Typing (A-B-O)

• Blood type A has antigen A on the surface of the
  cell and will agglutinate with blood type B.
• Blood type B has antigen B on the surface of the
  cell and will agglutinate with blood type A.
• Blood type AB has antigens A and B on the
  surface of the cells and will not agglutinate
  with either type A or B blood.
• Blood type O has neither antigen A nor B and will
  not agglutinate.

17
Antigen and Antibody
Antigens are large molecules (usually proteins)
on the surface of cells, viruses, fungi,
bacteria, and some non-living substances such as
toxins, chemicals, drugs, and foreign particles.
The immune system recognizes antigens and
produces antibodies that destroy substances
containing antigens.
18
BLOOD TYPES
19
Blood Typing Animation
Click here for animation and mini quiz
20
ABO Blood System
(Antigen)
Anti-A
Anti-B
Anti-A and Anti-B
none
http//gslc.genetics.utah.edu/units/basics/blood/t
ypes.cfm
21
Blood Typing and Cross-Reactions
Donor blood type is ____
Recipient blood type is ____
A
B
22
Blood Typing
Click for Blood typing simulation
23
Go to the following web site to play the blood
typing game

• First Go to How to play the game
• http//nobelprize.org/educational_games/medicine/l
  andsteiner/

24
ABO Blood Types

• Based on the presence (or absence) of antigen(s)
  on the red blood cell membrane
• There are 2 types of antigens that can be
  present,
• or
• A person can inherit one, both or none
• About 2 8 months after birth, you develop
  antibodies to the blood types you do not have.

A
B.
A (A antigen) B (B antigen) AB (both A B
antigen) O (no antigens)
25
Blood Transfusion
A person who is blood type A will have the ____
antigen on their RBC surface. In their
plasma, they will have anti- _____ antibodies. A
person who is blood type B will have the ____
antigen on their RBC surface. In their
plasma, they will have anti- _____ antibodies.
What will happen if a person with blood type B
donates blood to a person with blood type
A?
A
B
B
A
The B antibodies of the A plasma will attack and
destroy the B red blood cells, causing dangerous
and often fatal blood clotting.
26
Blood Groups
Antigen (on RBC)
Antibody (in serum)
Can Give Blood To
Can Get Blood From
Type
A
A
Anti-B
A, AB
O, A
B, AB
O , B
B
B
Anti-A
Neither Anti-A nor Anti-B
AB
A and B
AB
A, B, O, AB
Neither A nor B
A, B, O, AB
O
Anti-A and Anti-B
O
27
Population Distribution of Blood Types in the
U.S.
Type
Percent
O
43
A
42
B
12
AB
3
28
Blood Typing (Rh factor)

• Rh factor is determined by the presence of the D
  antigen.
• Rh positive people have the D antigen
• Rh negative people lack the D antigen

29
Secretors

• 80 of the population are secretors. Their
  blood-type antigens are found in high
  concentration in their body fluids such as
  saliva
• semen
• vaginal secretions
• gastric juice

30
Please Do Now
Please write at least 5 lines to explain this
cartoon in your composition book.
31
What is one thing you can give and still keep?

• Your word
• Hope
• Fingeprints
• Blood
• Name
• Life (pregnancy)
• Knowledge

32
Learning Objectives
Students should be able to

• List and describe forensic tests used to
  characterize a stain as blood.
• Understand the concept of antigen-antibody
  interactions and how it is applied to species
  identification and drug identification.

33
II. Immunoassay Techniques
34
Serology

• Describes a broad scope of laboratory tests that
  use specific antigen and serum antibody
  reactions.
• The concept of specific antigen-antibody
  reactions has been applied to immunoassay
  techniques for the detection of drugs of abuse in
  blood and urine.

35
Immunoassay

• The concept of antigen-antibody reaction is used
  in
• blood typing
• the detecting of drugs in blood and urine
• Thousands of individuals are subjected to
  urinalysis tests for the presence of drug-abuse.
  Such as
• Note Antibodies that react with drugs do NOT
  exist naturally but can be produced in animals
  such as rabbits.

• Military personnel
• Transportation industry employees
• Police and correction personnel
• Pre-employment drug screenings

36
Testing Urine for Drugs
One-step immunoassay for detection of drug abuse
in urine
Read Supreme Court Ruling
37
Immunoassay

• A number of immunological assay techniques are
  commercially available for detecting drugs
  through antigen-antibody reaction.
• Radioimmunoassay (RIA) uses drugs labeled with
  radioactive tags.
• Enzyme-multiplied immunoassay technique (EMIT) is
  uses an enzyme tag for detecting drugs in urine.
• Immunoassay is ONLY presumptive in nature and its
  result must be confirmed by additional testing.
  (not totally specific for any drug)

38
Radioimmunoassay (RIA)see p. 273 Fig. 8 - 3
uses drugs labeled with radioactive tags.
39
Enzyme-Multiplied Immunoassay Technique (EMIT)
see p. 274
used for detecting drugs in urine and fingernails
The method of screening for drug use in a
fingernail test is the five panel, Enzyme
Multiple Immunoassay Test (EMIT). Five drugs are
tested for with the qualitative detection of
cocaine, amphetamines / methamphetamines
(ecstasy), opiates (heroin, morphine),PCP (angels
dust,hog) and tetrahydrocannabinol (marijuana,
hashish) drug testing in urine. Results of a
fingernail sample will represent drug use that is
approximately 4-6 months from the time of
ingestion. Fingernails can be clipped, or, if
length does not allow, can be shaved in a safe
and pain-free sample collection.
40
Immunoassay

• Enzyme-multiplied immunoassay technique (EMIT) is
  used by toxicologists because of its
• speed for detecting drugs in urine and
• high sensitivity for detecting drugs in urine.
• In a typical EMIT analysis, antibodies that will
  bind to a specific drug are added to the
  subjects urine.

41
Antigen-Antibody Reaction(polyclonal antibodies)

• When an animal, such as a rabbit or mouse, is
  injected with an antigen its body will produce a
  series of different antibodies, all of which are
  designed to attack some particular site on the
  antigen of interest.
• This collection of antibodies is known as
  polyclonal antibodies.

42
Antigen-Antibody Reaction(monoclonal
antibodies)see p. 275

• A more uniform and specific collection of
  antibodies designed to combine with a single
  antigen site can be manufactured.
• Such antibodies are known as monoclonals.

43
III. Forensic Characterization of Bloodstains
IS IT BLOOD?
44
Unknown Stain at a Scene

• 3 Questions to be answered
• Is it blood?
• Is it human blood?
• Whose is it?
• Determine blood type, alcohol content, drugs
  present
• Determine the method(s) in which blood may have
  been deposited

The determination of blood is best made by means
of a preliminary color test.
45
Presumptive (Color) Tests for Blood Determination

• Kastle-Meyer color testa mixture of
  phenolphthalein and hydrogen peroxide the
  hemoglobin will cause the formation of a deep
  pink color if blood is present
• Hematest tabletreacts with the heme group in
  blood causing a green color
• Luminol testreaction with blood to produce light

46
Color Tests for Blood (Kastle - Meyer Tests)

• Replaced benzidine color test (after being
  identified as a carcinogen)
• a mixture of phenolphthalein and hydrogen
  peroxide
• the hemoglobin will cause the formation of a deep
  pink color if blood is present
• Not specific for blood - false positive given
  by
• Potatoes
• horseradish

Neither probably not encountered in criminal
situations
47
Color Tests for Blood (Hemastix strips or
Hematest tablets)

• reacts with the heme group in blood causing a
  green color

48
Presumptive (Color) Tests for Blood Determination
(Luminol)

• Reaction with blood to produce light
• Spray luminol on object
• Darken room
• Bloodstains react to produce a blue glow
  (luminescence)
• Used to check large areas
• Extremely sensitive
• Can detect bloodstains diluted up to ____________
  times
• Does NOT interfere with subsequent DNA testing

300,000
49
Microcrystalline Tests

• Depend on the addition of specific chemicals to
  the blood so that characteristic crystals will be
  formed
• ID of blood is made more specific if
  microcrystalline tests are done
• Less sensitive than color tests
• More susceptible to interference from
  contaminants present in the stain
• Two most popular tests are
• Takayama test and Teichmann test
• Click to view Takayama crystals

50
Precipitin Test (human vs. animal blood) see p.
279 Fig. 8 - 5

• Determines whether the stain is of human or
  animal origin
• Uses antisera normally derived from rabbits that
  have been injected with the blood of a known
  animal to determine the species origin of a
  questioned bloodstain.
• If human blood, then determine whose blood is it.

51
Precipitin Test see p. 279 Fig. 8 - 5

• blood is injected into a rabbit
• antibodies are formed
• the rabbits blood is extracted as an antiserum
• the antiserum is placed on sample blood.
• The sample will react with human proteins, if
  human blood is present.

52
Precipitin Test

• Why is this a good test for determining if it
  human blood?
• very sensitive
• requires only a small amount of blood
• Blood can be old and still give a positive result
• Bloodstains from 4000-5000 year old mummies
  have given positive results

53
Gel Diffusion

• antibodies and antigens diffuse or move towards
  one another on an agar plate.

54
Human vs Animal Blood
Microscopic observation
Frog Blood
Human Blood

• Larger nucleic red blood cells

nonnucleated red blood cells
55
IV. Bloodstain Patterns
Science of Murder Blood Spatter Video
56
More about Serology

• For additional information about blood evidence,
  and famous crimes that involves serology, check
  out Court TVs Crime Library at
• www.crimelibrary.com/criminal_mind/forensics/sero
  logy/1.html

57
People of Historical Significance Paul Kirk
(1902-1970)

• Professor of criminalistics and biochemistry at
  Berkeley in California
• He actively assisted law enforcement
  organizations from 1935 to 1967.
• His book, Crime Investigations, contained a
  chapter in which he discussed the application of
  blood stain pattern analysis to criminal
  investigations.
• Dr. Kirk analyzed the blood stain pattern photos
  from the Sam Sheppard case and was instrumental
  in Sheppards release at his second trial. Find
  out more about the case at CourtTvs crime
  library.

58
People in the News Herbert L. MacDonell

• Considered by many as the father of modern
  bloodstain pattern analysis.
• He is the director of the Lab of Forensic Science
  and founder of the Bloodstain Evidence Institute
  (1973) in Corning, NY.
• His work, Bloodstain Pattern Interpretation,
  helped to jump start this discipline.
• He has consulted on criminal cases in all 50
  states, in addition to testifying in the O.J.
  Simpson trial and in the assassination cases of
  Sen. Robert F. Kennedy and Dr. Martin Luther King
  Jr.

59
Blood Spatter Evidence

• A field of forensic investigation which deals
  with the physical properties of blood and and the
  patterns produced under different conditions as a
  result of various forces being applied to the
  blood. Blood, as a fluid, follows the laws of
  physics.

60
Blood Evidence

• Class evidence for blood would include blood
  type.
• If you can determine the DNA you would have
  individual evidence.
• Blood stain patterns are considered
  circumstantial evidence in a court room. Experts
  could argue many points including
• direction of travel,
• height of the perpetrator
• position of the victim
• left/right hand
• whether the body was moved, etc.

61
Blood Pattern Reconstruction

• Scene Pattern Reconstruction
• 1. Stain condition
• 2. Pattern
• 3. Distribution
• 4. Location
• 5. Directionality

• Lab Results Reconstruction
• 1. Genetic marker typing
• 2. Age Determination
• 3. Source Determination
• 4. Race Determination
• 5. Sex Determination

From Cracking Cases by Dr. Henry C. Lee
62
Questions Answered by Blood Spatter
Interpretation

• The distance between the target surface and the
  origin of blood
• The point(s) of origin of the blood
• Movement and direction of a person or an object
• The number of blows, shots, etc. causing the
  bloodshed and/or the dispersal of blood.
• Type and direction of impact that produced the
  bloodshed
• The position of the victim and/or object during
  bloodshed
• Movement of the victim and/or object after
  bloodshed

63
Bloodstain Patterns

• The crime scene investigator must remember that
  the location, distribution, and appearance of
  bloodstains and spatters may be useful for
  interpreting and reconstructing the events that
  produced the bleeding.
• Surface texture and the stains shape, size and
  location must be considered when determining the
  direction, dropping distance, and angle of impact
  of a bloodstain.

64
Blood Droplet Characteristics

• A blood droplet will remain spherical in space
  until it collides with a surface
• Once a blood droplet impacts a surface, a
  bloodstain is formed.
• A droplet falling from the same height, hitting
  the same surface at the same angle, will produce
  a stain with the same basic shape.
• How will the shape change as the height is
  increased or decreased?

click to view
65
Blood Droplet Volume

• A droplet contains approximately 0.05 cc of fluid
• Is not the same for all blood droplets, but is
  generally from 0.03 cc to 0.15 cc
• Is directly dependent upon the surface or orifice
  from which it originates
• The impact area is called the target.

66
Conditions Affecting Shape of Blood Droplet

• Size of the droplet
• Angle of impact
• Velocity at which the blood droplet left its
  origin
• Height
• Texture of the target surface
• On clean glass or plasticdroplet will have
  smooth outside edges
• On a rough surfacewill produce scalloping on the
  edges

Click to see
67
TARGET SURFACE TEXTURE
68
Target surface texture

• Bloodstains can occur on a variety of surfaces,
  such as carpet, wood, tile, wallpaper,
  clothing..
• The type of surface the blood strikes affects the
  amount of resulting spatter, including the size
  and appearance of the blood drops.

69
Target surface texture

• Blood droplets that strike a hard smooth surface,
  like a piece of glass, will have little or no
  distortion around the edge.

70
Target surface texture

• Blood droplets that strike linoleum flooring take
  on a slightly different appearance.
• Notice scalloping around the edge of the blood
  droplets.

71
Target surface texture

• Surfaces such as wood or concrete are distorted
  to a larger extent.
• Notice the spines and secondary spatter present.

72
Surface Texture

• In general, the harder and less porous the
  surface, the less spatter.
• The softer and more porous the surface, the more
  a blood drop will break apart.

73
Bloodstain Patterns(direction of travel)

• The pointed end of the blood stain faces the
  direction of travel.

Which way did the blood travel?
74
Determining angles of impact

• Blood droplets in freefall have the shape of a
  sphere.
• Droplets striking surfaces and leaving
  well-formed stains make it possible to determine
  the angle at which
• the droplet struck the surface.

75
Angle of Impact

• The more acute the angle of impact, the more
  elongated the stain.
• 90 degree angles are perfectly round drops with
  80 degree angles taking on a more elliptical
  shape.
• At about 30 degrees the stain will begin to
  produce a tail.
• The more acute the angle, the easier it is to
  determine the direction of travel.

76
Angle of Impact

• The shape of a blood drop
• Roundif it falls straight down at a 90 degree
  angle
• Ellipticalblood droplets elongate as the angle
  decreases from 90 to 0 degrees
• the angle can be determined by the following
  formula

77
Angle of Impact
78
Point of Convergence

• The point of convergence is the intersection of
  two bloodstain paths, where the stains come from
  opposite sides of the impact pattern

79
Area of Convergence(point of origin)

• The area of convergence is the box formed by the
  intersection of several stains from opposite
  sides of the impact pattern

80
Point or Area of Convergence?
81
Point of Convergence

• The location of the blood source can be
  determined by drawing lines from the various
  blood droplets to the point where they intersect.
  
• The area of convergence is the point of origin
  the spot where the blow occurred. It may be
  established at the scene with measurement of
  angles by use of strings.

82
Origin of blood
Image used with permission from Tom Bevel Ross
Gardner, June 2006.
83
Bloodstain pattern categories(based on the
mechanism that created them)

• Passive bloodstains are created when the force
  acting on the blood is gravity
• Projected bloodstains occur when some form of
  energy has been transferred to a blood source
• Transfer or contact is produced when an object
  with blood on it comes into contact with an
  object or a surface that does not have blood on
  it.

84
Bloodstain pattern categories
PASSIVE
PROJECTED
TRANSFER
85
Passive Bloodstains

• Drop(s) created or formed by the force of gravity
  acting alone.
• Drops
• Drip patterns
• Pools
• Clots

86
Projected Bloodstains

• Arise when a victim is subjected to blows that
  produce blood released at a force greater than
  gravity.
• 3 subcategories
• Impact spatter
• Cast-off stains
• Arterial gush
• The size, shape and number of resulting stains
  will depend, primarily on the amount of force
  utilized to strike the blood source.

87
Projected Bloodstains(impact spatter)

• Created when a blood source receives a blow or
  force resulting in the random dispersion of
  smaller drops of blood
• Subdivided into
• Low velocity spatter
• Medium velocity spatter
• High velocity spatter
• Back spatter
• Expiratory blood

88
Low Velocity Spatter

• Relatively large stains 4 mm in size and greater.
  Impact velocity up to 5 feet/sec (Blunt force)

89
Medium Velocity Spatter

• Preponderant stain size 1 to 4 mm size.
• Impact velocity of 5 to 25 feet/sec.

Image courtesy UWA PhD research student Mark
Reynolds.
90
High Velocity Spatter

• Preponderant stain size 1 mm in size and smaller.
  
• Mist like appearance.
• Impact velocity of 100 feet/sec and greater.
• (gun shot, high speed machinery)

91
Projected Bloodstains

• Back spatter blood directed back towards the
  source of energy or force that caused the spatter
• Expiratory blood blood that is blown out of the
  nose, mouth or a wound as a result of air
  pressure and/or air flow which is the propelling
  force.

92
Gunshot back forward spatter
Bloodstained foam held just above target surface.
Bullet passing L to R just above sheet
Back-spatter on entry
Forward spatter on exit
93
Which type of impact spatter?
Low velocity blood from the simulation of a
bleeding person walking or running. Note that
the blood drops point in the direction of travel
The high velocity gun shot wound leaves a
mist-like appearance.
Low and medium Velocity, lightly Magnified.
94
Projected Bloodstains(cast-off pattern)
Created when blood is released or thrown from a
blood-bearing object in motion
95
Arterial Spurting or Gushing

• Bloodstain pattern(s) resulting from blood
  exiting the body under pressure from a breached
  artery

96
Transfer (Contact) Bloodstains(gun, knife, hand,
foot)

• Occurs when an object contaminated with blood
  comes in contact with another surface
• Examples include
• Swipe pattern
• Wipe pattern
• Smudge
• Transfer pattern
• Blockage
• Simple direct contact

97
Swipe Pattern

• The transfer of blood from a moving source onto
  an unstained surface. Direction of travel may be
  determined by the feathered edge.

98
Wipe Pattern

• A bloodstain pattern created when an object moves
  through an existing stain, removing and/or
  altering its appearance.

99
Transfer (Contact) Bloodstains(Smudge)
100
Transfer Pattern

• A recognizable image of all or a portion of the
  original surface may be observed in the pattern.

101
Transfer Pattern

• A recognizable image of all or a portion of the
  original surface may be observed in the pattern.

102
Transfer Pattern

• A recognizable image of all or a portion of the
  original surface may be observed in the pattern.

103
Transfer (Contact) Bloodstains(Blockage)
104
Crime Scene Clean Up
View video clip about crime scene clean up
2nd video clip

• Please DO NOW
• What do you think about the crime scene clean up
  business? Be sure to include something specific
  from the video clip.
• Do you think you could do this?
• Why or why not?

105
Bloodstain Terminology

• Angle of impactangle at which blood strikes a
  target surface.
• Bloodstain transferwhen a bloody object comes
  into contact with a surface and leaves a
  patterned blood image on the surface
• Backspatterblood that is directed back toward
  the source of energy
• Cast-offblood that is thrown from an object in
  motion

106
Bloodstain Terminology

• Contact stainbloodstains caused by contact
  between a wet blood-bearing surface and a second
  surface which may or may not have blood on it
• Transferan image is recognizable and may be
  identifiable with a particular object
• Swipewet blood is transferred to a surface which
  did not have blood on it
• Wipea non-blood bearing object moves through a
  wet bloodstain, altering the appearance of the
  original stain

107
Bloodstain Terminology

• Directionalityrelates to the direction a drop of
  blood traveled in space from its point of origin
• Terminal velocitythe greatest speed to which a
  free falling drop of blood can accelerate in air.
  It is dependent upon the acceleration of gravity
  and the friction of the air against the
  bloodapproximately 25.1 feet/second.
• High velocitygreater than 25 feet per second,
  usually 100 feet per second gives a fine mist
  appearance
• Medium velocity5 to 25 feet per second
• Low velocityup to 5 feet per second

108
Tips for the Chp. 8 Test

• Multiple Choice
• True / False
• Short Answers
• What can blood stains tell you?
• Blood Types Typing
• (antigen antibody)
• Blood spatter analysis
• Impact angle, shape, size,etc.
• Blood tests (precipitin, etc.)
• Blood - general information

• What 3 questions are asked at a bloody crime
  scene?
• Monoclonal antibodies vs. polyclonal antibodies
• Paternity testing
• Why might there not be any sperm in semen?
• Sexual assault cases (what is collected?)

109
Learning Objectives
Students should be able to

• Contrast chromosomes and genes.
• Use the Punnett square to determine the genotypes
  and phenotypes of offspring.

Fathers genotype
O O
A
Mothers genotype
B
110
V. Principles of Heredity
All of the antigens, polymorphic enzymes and
proteins previously discussed are genetically
controlled traits.
111
Principles of Heredity

• Genes (basic unit of heredity) are located on
  chromosomes.
• The position a gene occupies on a chromosome is
  its locus.
• Alleles are alternative forms of genes that
  influence a given characteristics (such as blood
  type)
• Each cell (except for eggs and sperm) have _____
  chromosomes.

46
Click for Web Extra 8.2
Click for Web Extra 8.3
112
Gene Pair

• A gene pair is made up of two alleles.
• Homozygous gene pair of 2 similar alleles
• Ex. AA or BB or OO blood types
• Heterozygous gene pair of 2 different alleles
• Ex. AO or BO or AB blood types
• One gene can be dominant over the other in a gene
  pair of different alleles.
• Ex. A and B are dominant over O in blood
  types.
• ( O is the recessive gene)
• In AB blood type, the genes are codominant.

Click for Web Extra 8.5
113
Genotype and Phenotype

• Phenotype a persons outward appearance
• Ex. You have B type blood
• Genotype a persons genetic makeup for a trait
• Ex. You have BB or BO type blood.
• No blood test can determine your genotype.
• By studying the family history of an individual
  you may be able to determine their genotype

Click for Web Extra 8.4
114
Punnett Squares and Paternity
PossibleFathers genotype
O O
What are the possible blood types for their
children?
AO
A
AO
Sallys genotype
BO
BO
B
Could he be the father of her AB baby boy? Why or
why not?
115
Paternity Testing

• Disputed paternity cases are normally encountered
  in civil, not criminal courts.
• Genotyping of blood antigens (factors) can be
  useful in determining paternity by ruling out a
  suspected father.
• A-B-O grouping
• HLA (human leukocyte antigen)- antigens on WBC
• If it cant exclude a suspect than its better
  than 90 that hes the father
• DNA testing
• Can determine with better than 99 that he is the
  father

116
Please Do NowExplain this cartoon.
117
Learning Objectives
Students should be able to

• List the laboratory tests necessary to
  characterize seminal stains.
• Explain how suspect blood and semen stains are
  properly preserved for laboratory examination.

UV light makes seminal fluids glow brightly
118
VI. Forensic Characterization of Semen

• Two step process for the examination of seminal
  stains
• Locate the stain
• Test the stain to prove its semen

119
Testing for Seminal Stains

• Seminal stains may be visible on fabric due to
  their stiff, crusty appearance.
• Acid phosphatase test is the best way to locate
  and characterize seminal stains.
• Once sample is proven to be semen, the next step
  is to associate the semen as closely as possible
  with an individual

120
Acid Phosphatase Test

• Acid phosphatase is an enzyme secreted by the
  prostate gland into seminal fluid.
• Concentration is 400X more in seminal fluid than
  in any other body fluid.
• A reaction time of less than 30 seconds is a
  strong indication of semen.

121
Testing Seminal Stains for acid
phosphatase
Moisten Stain Collect Stain Open Test
Strip Purple semen Activate stain Rub
stain with Test stain by Semen
is present with water. provided cotton
rubbing the if it turns purple
swab. moistened swab
immediately. onto test strip.
(lt 30 seconds)
122
Microscopic Examination of Semen

• Semen is unequivocally identified by the presence
  of spermatozoa.
• Usually easy to locate sperm in semen
• Reasons why sperm might not be found
• Sperm bind tightly to cloth material
• Sperm are extremely brittle when dry and easily
  disintegrate when washed or rubbed against
  another object
• Oligospermia lows sperm count
• Aspermia no sperm in seminal fluid

123
Seminal constituents sperm

• Live (motile) sperm generally survive for up to 4
  - 6 hours in the vaginal cavity
• Vaginal smear must be examined microscopically
  immediately after it is taken from the victim
• Nonmotile sperm may be found up to 3 days in the
  vaginal cavity (occasionally up to 6 days later)
• Intact sperm (sperm with tail) are not normally
  found 16 hours after intercourse (but have been
  found 72 hours later)

124
Seminal constituents

• Finding acid phosphatase decreases with time
  after intercourse
• Little chance of identifying it after 48 hours
• Need to know if voluntary sexual activity
  occurred before the assault
• p30 is NOT normally found in the vaginal cavity
  beyond 24 hours after the assault

125
Prostate Specific Antigen (PSA or
p30)

• Positive acid phosphatase test but cant find any
  sperm how can you prove unequivocally that its
  semen?
• By use of p30 (prostate specific antigen, PSA)
• Antigen antibody reaction
• see p. 293 Figure 8-17 and Figure 8-18

126
Learning Objectives
Students should be able to

• Describe the proper collection of physical
  evidence in a rape investigation.

127
VII. Collection of Rape Evidence

• Seminal constituents on a rape victim indicate
  that sexual intercourse occurred BUT their
  absence does not necessarily mean that a rape did
  not occur.
• Bruises and bleeding tend to confirm a violent
  assault occurred
• Physical evidence of rape may include semen,
  blood, hairs and fibers

128
How to protect rape evidence

• Outer garments and undergarments carefully
  removed and packaged in separate paper bags.
  WHY?
• Dont fold an article through a seminal stain as
  it may damage the sample.
• Latex gloves must be worn when collecting samples

129
How to protect rape evidence(the victim)

• The rape victim must undergo a medical
  examination as soon as possible after the assault
• Use an evidence collection kit
• see p. 295 figure 8-19a and figure 8-19b and p.
  296 Figure 8-19c

130
Physical evidence to be collected from scene/
victim

1. Pubic combings
2. Pubic hair standard/ reference samples
3. External genital dry-skin areas
4. Vaginal swabs and smear
5. Cervix swabs
6. Rectal swabs and smear
7. Oral swabs and smear

131
Physical evidence to be collected from scene/
victim

• Head hairs
• Blood sample (for DNA)
• Fingernail scrapings
• All clothing
• Urine specimen
• Check for Rohypnol, GHB, etc.

132
Physical evidence to be collected from suspect

1. All clothing
2. Pubic hair combings
3. Pulled hair and pubic hair standard/reference
   samples
4. Penile swab (within 24 hours of assault)
5. Blood sample or buccal swab (for DNA)

133
DNA Fingerprinting Contd

• This will therefore produce a unique banding
  pattern following a gel electrophoresis.
• This test is highly accurate, and the probability
  of another individual possessing an identical
  banding pattern is estimated as around
  114,000,000,000.

134
DNA Fingerprinting