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Forensic Serology
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Serology-study of biological fluids involving specific antigen and serum antibody
reactions
• 1901-Karl Landsteiner-ABO blood type system• Blood types based on presence of
agglutinogens (agglutinating antigens) on surface of red blood cells (erythrocytes)
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• Proteins that stimulate production of antibodies
• On surface of erythrocytes• Types- A and B• Antibodies-produced against antigens not
present• Ex. Type A blood has A antigens, but B
antibodies
Antigens
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Blood Types
Blood Type
Antigen Antibodies in plasma
Can give to
Can receive from
A A Anti B A,AB O, A
B B Anti A B,AB O.B
AB AB No A or B AB O,A,B,AB
O Neither A nor B
A and B O,A,B,AB
O
http://nobelprize.org/educational_games/medicine/landsteiner/
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Frequency of Blood Types
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Blood Factors
• If mixed-blood agglutinates (solidifies) -death
• Rh factor (antigen D)• More than 100 blood factors
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Blood
1. Cells-Red blood cells
(Erythrocytes)Platelets (Thrombocytes)White blood cells
(Leukocytes)
2. Enzymes3. Proteins4. Fluid5. Inorganic
substances (gases)
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Blood Plasma
• Liquid part of blood • Serum (liquid that separates from blood)
and proteins in water• 55% of blood volume• Fibrin-blood clotting protein
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Erythrocytes
• Red blood cells• Transport oxygen and carbon
dioxide• Has hemoglobin-blood protein
carries oxygen in blood• Most numerous blood cells
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Leukocytes (White Blood Cells)
• Monocytes• Lymphocytes• Neutrophils• Part of Immune System• Fight infections
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Platelets• Thrombocytes• Involved in blood clotting
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Antiserum
• Serum that contains antibodies (proteins that destroy specific antigens ;immunoglobins)
• Used in determining blood types• If antiserum causes agglutination,
then the blood type can be determined
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Bloodstain Characteristics
• As blood dries-harder to type• Erythrocytes lyse, or break open, upon
drying• No cells in dried bloodstain-antigens will
be present
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• Determine if it is blood• Determine species• If human blood-determine if it can be
individualized• Blood is class evidence
Guideline for Examining Blood
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1. Is it blood?2. From what species did the blood originate?3. If the blood is human, how closely can it be matched to one individual?
Forensic Characterizations of Bloodstains
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Is it Blood?Kastle-Meyer Color Test
• Phenolphthalein• Hemoglobin-peroxidase-like activity• Peroxidases-speed up oxidation of
compounds by peroxides• Blood, phenolphthalein, and hydrogen
peroxide-hemoglobin is deep pink
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1.) Hydrogen peroxide2.) Heme has peroxidase-like activity-breaks up
peroxides.H2O2 decomposed to H2O and oxygen3.) Oxygen combine with the KM, phenolphthalein color-
indicator4.) When KM solution is prepared-boiled remove oxygen-
When it turns colorless-shows KM had most of its oxygen removed and is now "oxygen hungry". So... now we have the makings of a kind of love story: the KM wants oxygen, and there's a lonely oxygen radical floating around, un-spoken for
5.) Pink color: occurs when oxygen radical "hooks up" with KM reagent-oxidation of KM and causes it to turn pink.
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Luminol TestChemiluminescence of luminolThe most sensitive chemical test that is capable of presumptively detecting bloodstains diluted up to 300,000 times
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• Luminol (C8H7O3N3) mixed with hydrogen peroxide and hydroxide
• If hemoglobin and luminol interact-iron in hemoglobin catalyzes rxn b/w H2O2 and luminol
• Redox rxn-luminol loses N and H and gains O-produce 3-aminophthalate
• 3-aminophthalate in energized state –e-s in oxygen are boosted to higher orbitals-e-s then fall back to lower energy level-emit extra energy as light photons
• http://www.howstuffworks.com/framed.htm?parent=luminol.htm&url=http://www.shsu.edu/%7Echm_tgc/JPPdir/JPP1999/
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Immunoassay TechniquesPresumptive test for detection of drugs in blood and urine
Antibodies reacting with drugs don’t occur naturally
Produced in animals by combining drug with a protein and injecting into animal
Drug-protein complex acts as antigen stimulating animal to produce antibodies
The recovered blood serum of animal contains specific antibodies to the drug-This is human antiserum
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EMIT- Enzyme Multiplied Immunoassay Technique-drug screening in urine and blood-add antibodies to blood or urine, then add chemically labeled drug- will compete with drugs in urine or blood (if present) for antibody binding sites. The number of remaining unbound labeled drug-proportional to drug’s concentration in urine.ex. Methadone-add methadone antibodies and chemically-labeled methadone to urine- ex. THC tetrahydrocannabinol- metabolite-THC-9-carboxylic acid-antibodies are prepared against this metabolite-one-millionth of a gram-can detect up to 10 days after use
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Steps required to produce monoclonal antibodies
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Polyclonal vs. Monoclonal Antibodies
• Polyclonal antibodies-series of antibodies which can attack some site on antigen-vary in specificity and binding abilities
• Monoclonal antibodies-identical antibodies that interact with one site on an antigen
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Monoclonal antibodies
• Immunoassay tests for abused drugs made with monoclonal antibodies
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Precipitin Test
• Test to determine species of blood• Precipitin-an antibody that reats with its
corresponding antigen to form a precipitate
• Animal blood injected with human blood-antibodies form that react with human blood to neutralize its presence
• Antibodies can be recovered in serum• Serum is human anti-serum
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•Precipitin tests•Blood is placed on antiserum in tube•Blood will react with anitbodies in antiserum•Leaves precipitin band
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Gel Diffusion
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Blood Enzymes and Proteins
• Enzymes-proteins that regulate chemical reactions
• Polymorphic enzymes-enzymes of many forms
• Ex. PGM-Phosphoglucomutase-ten types of PGM
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Secretors
• 80 % of individuals are secretors• Secretors-antigens in most body fluids
(saliva, semen) and blood
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Bloodstains and Probability
• Antigens, enzymes, and proteins occur independently
• Probability-product of their distributions-Rule of Multiplication
• Ex. If bloodstain is type A, it originated from 42% of population. If it has PGM 1-origin narrowed to 24%
• 42% X 58%
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Study performed by Herbert MacDonell:
1. Surface texture- harder and less porous surface-less spatter
2. When blood drop hits hard surface-breaks up on impact-casting off smaller droplets
3. Impact angle of blood-Inverse Sin W/LW=width of stainL=length of stain
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Area of convergence on a two-dimensional plane. Convergence represents the area from which the stains emanated.
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http://www.nfstc.org/links/animations/images/blood%20spatters.swf
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Categories of BloodstainsPASSIVE
TRANSFER
PROJECTED
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By measuring the length and width of a bloodstain, the impact angle can be calculated
SIN < = Width (a) 1.5cm Length (c) 3.0cm Width (a) 1.5cm = SIN < Length (c) 3.0cm
0.5 = SIN <
< = 30 degrees
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Collection of Wet Blood
• Collected and refrigerated• Insufficient amount-air-dry • No plastic containers• Each stained article-packaged in paper
bag or cardboard box
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Drying Cabinets in Crime Lab
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Collection of Dried Bloodstains
• Stained sample packaged• Sample of unstained surface removed-
substrate control to confirm test results from stain and not substrate on which it was deposited
• Absorb blood onto damp swab
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Blood Collection
• Bloodstains compared to blood specimens from victims and suspects
• Collect 5cc from individuals-add anticoagulant EDTA (ethylenediamine tetraacetic acid)-inhibits enzymes from degrading DNA
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Heredity of Blood Types
• Genes-basic unit of heredity-located in DNA
• Chromosomes-structures made of DNA• Humans-46 chromosomes in somatic cells• 23 chromosomes in gametes• Zygote-fertilized egg• Alleles-forms of genes
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Multiple Alleles of Blood Types
• Homozygous-gene pair made of two similar genes-ex. AA and OO
• Heterozygous-gene pair made up of two different genes-AO
• A and B-dominant genes• O-recessive gene• AB-codominant
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Genotype and Phenotype• Genotype-pair of alleles-no laboratory test to
determine genotype ex. AA or AO• Phenotype-physical characteristic-ex. Type A
blood• Punnett Squares used to determine genotypes• Used in Paternity Testing
Mother
Fathero
A
B
o
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Forensic Characterization of Semen
• Each Male Ejaculate-2.5-6 mL of seminal fluid
• Each mL-100 million or more spermatozoa
• Acid-Phosphatase Color Test-enzyme secreted by prostate gland
• Oligospermia-low sperm count• Aspermia-no spermatozoa in seminal fluid
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• If semen is detected, but contains no spermatozoa, how do you prove presence of semen?
• Protein called p30-unique to seminal plasma