FORENSIC SEROLOGY

2. FORENSIC SEROLOGY Chapter 8

3. Nature of Blood • The word blood refers to a highly complex mixture of cells, enzymes, proteins, and inorganic substances. • Plasma, which is the fluid portion of blood, is composed principally of water. • Red blood cells (erythrocytes), white blood cells (leukocytes), and platelets are the solid materials suspended in plasma. • Antigens, usually proteins, are located on the surface of red blood cells and are responsible for blood-type characteristics.

4. Blood Typing • More than 15 blood antigen systems have been identified, but the A-B-O and Rh systems are the most important. • An individual that is type A has A antigens on his/her red blood cells, type B has B antigens, AB has both A and B antigens, and type O has neither A nor B antigens. • Rh factor is determined by the presence of another antigen, the D antigen. • People having the D antigen are Rh positive; those not having the antigen are Rh negative. • For every antigen there is a specific antibody that will react with it to form clumps known as agglutination. • Thus, if serum containing anti-B is added to red blood cells carrying B antigen, they will immediately react.

5. Serology • The term serology is used to describe a broad scope of laboratory tests that use specific antigen and serum antibody reactions. • 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. • The concept of specific antigen–antibody reactions has been applied to immunoassay techniques for the detection of drugs of abuse in blood and urine.

6. Immunoassay • A number of immunological assay techniques are commercially available for detecting drugs through antigen-antibody reaction. • One such technique, the enzyme-multiplied immunoassay technique (EMIT), is used by toxicologists because of its speed 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 subject’s urine. • Other immunoassay procedures are also available, such as radioimmunoassay (RIA), which uses drugs labeled with radioactive tags.

7. Antigen-Antibody Reaction • 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. • Alternately, 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.

8. Forensics of Blood • The criminalist must be prepared to answer the following questions when examining dried blood: 1. Is it blood? 2. From what species did the blood originate? 3. If the blood is of human origin, how closely can it be associated to a particular individual? • The determination of blood is best made by means of a preliminary color test.

9. The Tests • A positive result from the Kastle-Meyer color test is highly indicative of blood. • Hemoglobin causes a deep pink color. • Alternatively, the luminol test is used to search out trace amounts of blood located at crime scenes. • Produces light (luminescence) in a darkened area. • Microcrystalline tests, such as the Takayama and Teichmann tests, depend on the addition of specific chemicals to the blood so that characteristic crystals will be formed.

10. The Tests • Once the stain has been characterized as blood, the precipitin test will determine whether the stain is of human or animal origin. • The precipitin test 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. • Once it has been determined that the bloodstain is of human origin, an effort must be made to associate or dissociate the stain with a particular individual. • DNA analysis has allowed forensic scientists to associate blood to a single individual.

11. Stain Patterns of Blood • 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 stain’s shape, size, and location must be considered when determining the direction, dropping distance, and angle of impact of a bloodstain.

12. Stain Patterns of Blood • Surface texture is of paramount importance. In general, the harder and less porous the surface, the less spatter results. • The direction of travel of blood striking an object may be discerned because the pointed end of a bloodstain always faces its direction of travel. • The impact angle of blood on a flat surface can be determined by measuring the degree of circular distortion. At right angles the blood drop is circular, as the angle decreases, the stain becomes elongated. • 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 origin point.

13. Heredity and Paternity • The transmission of hereditary material is accomplished by means of microscopic units called genes, located on chromosomes. • Alternative forms of genes that influence a given characteristic (such as eye color or blood type) are known as alleles. • Paternity testing has historically involved the A-B-O blood typing system, along with blood factors other than A-B-O. • Currently, paternity testing has implemented DNA test procedures that can raise the odds of establishing paternity beyond 99 percent.

14. Testing for Seminal Stains • Many of the cases sent to a forensic laboratory involve sexual offenses, making it necessary to examine exhibits for the presence of seminal stains. • The best way to locate and at the same time characterize a seminal stain is to perform the acid phosphatase (an enzyme secreted into seminal fluid) color test. • A purple color indicates acid phosphatase enzyme. • Semen can be unequivocally identified by either the presence of spermatozoa or of p30, a protein unique to seminal plasma. • Forensic scientists can successfully link seminal material to an individual by DNA typing.

15. Rape Evidence • The rape victim must undergo a medical examination as soon as possible after the assault. • At that time the appropriate items of physical evidence including clothing, hairs, and vaginal and rectal swabs can be collected for subsequent laboratory examination. • All outer and undergarments should be carefully removed and packaged separately in paper (not plastic) bags. • Bedding, or the object upon which the assault took place, may also be carefully collected.

16. Rape Evidence • If a suspect is apprehended within 24 hours of the assault, it may be possible to detect the victim’s DNA on the male’s underwear or on a penile swab of the suspect. • Items routinely collected from the suspect include all clothing, pubic hair, head hair, penile swab, and a blood sample or buccal swab for DNA typing. • The forceful physical contact between victim and assailant may result in a transfer of such physical evidence of blood, semen, saliva, hairs, and fibers.

17. The word blood refers to a highly complex mixture of cells, enzymes, proteins, and inorganic substances. • Plasma, which is the fluid portion of blood, is composed principally of water.

18. Red blood cells (erythrocytes), white blood cells (leukocytes), and platelets are the solid materials suspended in plasma. • Antigens, usually proteins, are located on the surface of red blood cells and are responsible for blood-type characteristics.

19. Blood Typing • More than 15 blood antigen systems have been identified, but the A-B-O and Rh systems are the most important. • An individual that is type A has A antigens on his/her red blood cells, type B has B antigens, AB has both A and B antigens, and type O has neither A nor B antigens

20. People having the D antigen are Rh positive; those not having the antigen are Rh negative. • For every antigen there is a specific antibody that will react with it to form clumps known as agglutination. • Thus, if serum containing anti-B is added to red blood cells carrying B antigen, they will immediately react.

21. Serology • The term serology is used to describe a broad scope of laboratory tests that use specific antigen and serum antibody reactions. • 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.

22. The concept of specific antigen–antibody reactions has been applied to immunoassay techniques for the detection of drugs of abuse in blood and urine.

23. Immunoassay • A number of immunological assay techniques are commercially available for detecting drugs through antigen-antibody reaction. • One such technique, the enzyme-multiplied immunoassay technique (EMIT), is used by toxicologists because of its speed and high sensitivity for detecting drugs in urine.

24. In a typical EMIT analysis, antibodies that will bind to a specific drug are added to the subject’s urine. • Other immunoassay procedures are also available, such as radioimmunoassay (RIA), which uses drugs labeled with radioactive tags

25. Antigen-Antibody Reaction • 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. • Alternately, 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

26. Forensics of Blood • The criminalist must be prepared to answer the following questions when examining dried blood: • 1. Is it blood? • 2. From what species did the blood originate? • 3. If the blood is of human origin, how closely can it be associated to a particular individual? • The determination of blood is best made by means of a preliminary color test.

27. The Tests • A positive result from the Kastle-Meyer color test is highly indicative of blood. Hemoglobin causes a deep pink color. • Alternatively, the luminol test is used to search out trace amounts of blood located at crime scenes. Produces light (luminescence) in a darkened area. • Microcrystalline tests, such as the Takayama and Teichmann tests, depend on the addition of specific chemicals to the blood so that characteristic crystals will be formed. • Once the stain has been characterized as blood, the precipitin test will determine whether the stain is of human or animal origin. • The precipitin test 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.

28. Stain Patterns of Blood • 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 stain’s shape, size, and location must be considered when determining the direction, dropping distance, and angle of impact of a bloodstain. • Surface texture is very important In general, the harder and less porous the surface, the less spatter results.

29. The direction of travel of blood striking an object may be discerned because the pointed end of a bloodstain always faces its direction of travel. • The impact angle of blood on a flat surface can be determined by measuring the degree of circular distortion. At right angles the blood drop is circular, as the angle decreases, the stain becomes elongated. • 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 origin point.

30. Heredity and Paternity • The transmission of hereditary material is accomplished by means of microscopic units called genes, located on chromosomes. Chromosomes are condensed DNA • Alternative forms of genes that influence a given characteristic (such as eye color or blood type) are known as alleles. Alleles are found at loci of each chromosome • Paternity testing has historically involved the A-B-O blood typing system, along with blood factors other than A-B-O. • Currently, paternity testing has implemented DNA test procedures that can raise the odds of establishing paternity beyond 99 percent. • Punnett squares for blood type

31. Testing for Seminal Stains • The best way to locate and at the same time characterize a seminal stain is to perform the acid phosphatase (an enzyme secreted into seminal fluid) color test. A purple color indicates acid phosphatase enzyme. • Semen can be unequivocally identified by either the presence of spermatozoa or of p30, a protein unique to seminal plasma. • Forensic scientists can successfully link seminal material to an individual by DNA

32. Rape Evidence • The rape victim must undergo a medical examination as soon as possible after the assault. • At that time the appropriate items of physical evidence including clothing, hairs, and vaginal and rectal swabs can be collected for subsequent laboratory examination. • All outer and undergarments should be carefully removed and packaged separately in paper (not plastic) bags. • Bedding, or the object upon which the assault took place, may also be carefully collected.

35. Bedding, or the object upon which the assault took place, may also be carefully collected. • If a suspect is apprehended within 24 hours of the assault, it may be possible to detect the victim’s DNA on the male’s underwear or on a penile swab of the suspect. • Items routinely collected from the suspect include all clothing, pubic hair, head hair, penile swab, and a blood sample or buccal swab for DNA typing. • The forceful physical contact between victim and assailant may result in a transfer of such physical evidence of blood, semen, saliva, hairs, and fibers.

36. Passive bloodstains • Passive bloodstains are those stains created by the force of gravity. • Passive Drops - Bloodstain drop(s) created or formed by the force of gravity acting alone. • Drip Pattern - A bloodstain pattern which results from blood dripping into blood. • Flow Pattern - A change in the shape and direction of a bloodstain due to the influence of gravity or movement of the object. • Pool Pattern - A bloodstain pattern formed when a source of blood is stationary for a period of time.

37. Projected bloodstains • A projected stain occurs when some form of energy has been transferred to a blood source. • Low Velocity Impact Spatter (LVIS) - A bloodstain pattern that is caused by a low velocity impact\force to a blood source. • Medium Velocity Impact Spatter (MVIS) - A bloodstain pattern caused by a medium velocity impact\force to a blood source. A beating typically causes this type of spatter. • High Velocity Impact Spatter (HVIS) - A bloodstain pattern caused by a high velocity impact\force to a blood source such as that produced by gunshot or high-speed machinery. • Cast-Off Pattern - A bloodstain pattern created when blood is released or thrown from a blood-bearing object in motion.

38. Projected bloodstains • Arterial Spurting (OR Gushing) Pattern - Bloodstain pattern(s) resulting from blood exiting the body under pressure from a breached artery. • 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.

39. Transfer/Contact bloodstains • A transfer or contact stain is produced when an object with blood comes in contact with an object or surface that does not have blood. It may be possible to discern the object that left the blood impression. • Wipe Pattern - A bloodstain pattern created when an object moves through an existing stain, removing and/or altering its appearance. • Swipe Pattern - The transfer of blood from a moving source onto an unstained surface. Direction of travel may be determined by the feathered edge.

40. "Velocity" impact stains • Contrary to what the name states, the terms low-, medium-, and high-velocity impact spatter do not describe the velocity of the blood droplets as they fly through the air. The variation in the "velocity" is meant to describe the amount of energy transferred to a blood source in order to create the stains. Velocity is a speed (m/s) with a direction

41. Low velocity impact spatter • Low velocity impact spatter (LVIS) is generally produced when objects traveling less than 1.5 m/s come in contact with a blood source. The preponderance of stains is generally larger than 3 mm in diameter.

42. Medium velocity impact spatter • Medium velocity impact spatter (MVIS) is generally produced when objects traveling between 1.5 m/s and 7.5 m/s come in contact with a blood source. The preponderance of stains is generally between 1 mm and 3 mm in diameter. Mechanisms that could produce this type of pattern include blunt force trauma or cutting/stabbing actions.

43. High velocity impact spatter • High velocity impact spatter (HVIS) is generally produced when objects traveling greater than 30 m/s come in contact with a blood source. The preponderance of stains is generally smaller than 1 mm in diameter. This pattern often has a mist-like appearance. High velocity patterns may be created by gunshots or explosives, but may also be caused by industrial machinery, coughing, or sneezing.

46. angle of impact = arcsin (opposite side/hypotenuse) • Here's what an analyst has to do to for this to work: • Measure the length and width of the splatter. • Divide the width of the splatter by its length. • Determine the arcsin of that number, typically using a calculator with an arcsin function. • A drop of blood that fell perfectly vertically, or at a 90-degree angle, will be round. As the angle of impact increases, the drop of blood gets longer and develops a "tail." This tail points in the direction that the drop traveled, but its length isn't part of the measurements.