BLOOD AND STAIN

The test for the identification of blood is employed as an important part of the routine investigation in
many cases of violent death. The specimen usually submitted is fresh blood or fluid blood, dried blood and
clotted blood. Very often it is brought to the laboratory as dried blood or brown stain on weapons, clothing or
other objects.

IMPORTANCE OF THE STUDY OF BLOOD

1. As circumstantial or corroborative evidence against or in favor of the perpetrator of the crime.
2. In case of disputed parentage.
3. Determination of the cause of death and the length of time the victim survived the attack.
4 Determination of the direction of escape of the victims or the assailant.
5. Determination of the origin of the flow of blood.
6. Determination of the approximate time the crime was committed.

BLOOD
• Has been called the circulating tissue of the body
• Made up of formed elements suspended in a liquid called plasma
• Highly complex mixtures of cells, enzymes, proteins and inorganic substances
• About 6 quarts of blood in an average size man.
• It is opaque
• Slightly alkaline (normal pH is 7.35-7.45)

COMPOSITION OF BLOOD
1. 35% formed elements
a) Red blood cells or erythrocytes- contains hemoglobin and carries oxygen to various tissue in the
body.
b) White blood cells or leukocytes- defends the body from invading microorganism and help fight
infection.
c) Blood platelets or thrombocytes-produced by the bone marrow and responsible for proper clotting of
blood.
2. 65% plasma-fluid portion of the blood where cells are suspended
a) 90% water
b) 10% solid-largely protein in nature and consists of the following:
Albumen - the most abundant protein of the blood globulins - important role in the immune
mechanism of the body.
Fibrinogen-soluble precursors of fibrin which forms blood clot

PROBLEMS IN THE STUDY OF BLOOD

1. Where blood has to be searched?
2. Collection, preservation and transportation of specimen suspected to contain blood.
3. Does the stain contain blood or another substance?

CASES WHERE BLOOD CAN BE USED AS EVIDENCE

1 Stabbing
2 Mauling
3 Hit and run
4. Rape
5. Paternity cases
6. Robbery/burglary

COLLECTION OF BLOOD STAIN

In collecting blood stain, use a cotton swab wet with Normal Saline Solution (NSS), rub until you
absorb all blood in one place. Use different swab for different blood stain in different places. Do not destroy
furniture or bring the whole linen where the blood stain is found. In cases involving blood, you need a standard
sample from the victim (in case it is a cadaver) and also from the suspect. Blood stain is a needed sample not
decomposed blood. Do not let your sample to decomposed

CHRONOLOGICAL TEST FOR BLOOD

1. Preliminary test
2. Confirmatory test
3. Precipitin test
4. Blood grouping

PRELIMINARY TEST - determine whether the stain contains blood or another substance.
PRELIMINARY TEST FOR BLOOD
1. Benzidine test
 Very sensitive test that can be applied to minute stain
 Very delicate test and detect blood when present in a dilution of 1;300,000 parts
 It use was discontinued because it was known to be carcinogenic
 Can be used to very old stain or decomposed stain with all sorts of contamination.
Positive result intense blue color produced immediately
2. Guaiacum test
 Fairly delicate test showing presence of blood in a solution of 1;50,000 dilution.
 May not react to very old stain
 Limitation of the test reacts with saliva, pus, bile, milk, rust, iron, salt, cheese, potatoes, perspiration and
other oxidizing substances.
Positive result. Beautiful blue color that appears immediately
3. Phenolpthalein test (Kustle-Meyer test)
 Alternative test to benzidine test.
 Can detect blood in a dilution of 1:80,000,000 parts.
  Positive result is highly indicative of blood
 The test also give positive results for copper salts, potatoes and horseradish.
Positive result: rose color develops/deep pink
4. Leucomalachite Green test
 Recommended by Adler in 1904
 Not as sensitive as the Benzidine test
Positive Result malachite green or bluish green
5. Luminol test-important presumptive identification for blood
 Very sensitive reagent that undergoes oxidation by hydrogen peroxide in alkaline solution in the
presence of the heme fraction of hemoglobin
 Reaction of luminal with blood results in the production of light rather than color
 Extremely sensitive test(diluted up to 10,000 times
Positive result: luminescence or emission of light
6. Fluorescein test - like Luminol, emits light when exposed to an oxidant and heme. Unlike Luminol, however,
fluorescein undergoes fluorescence rather than chemiluminescence. It is applied to a suspected bloodstain along
with hydrogen peroxide. A strong, short-wave light is then used to induce fluorescence
LOCATING BLOOD ON OBJECTS
• The fact that a stain is dark red or black may mean that it is blood. Sometimes these stains are very small or
are on dark surfaces that mask their presence. In some cases, blood has been washed off the surface. Some tests.
• These also serve as preliminary tests for blood. The two major tests which can help locate bloodstains are
Luminol and fluorescein.

CONFIRMATORY TEST - determines whether bloodstain really contains blood.

CONFIRMATORY TEST FOR BLOOD
• The actual proof that a stain is blood is establishing the presence of the characteristic pigment, hemoglobin or
one of its derivatives
• Hemoglobin is the red coloring matter of the RBC of the blood
• The 3 confirmatory test for blood
1. Microscopic test
2 Microchemical test or microcrystalline test
3. Spectroscopic test
1. Microscopic test
 Useful in the demonstration and mensuration of blood cells
 Use to differentiate between mammalian, avian, piscine and reptilian blood
2. Microchemical test and Microcrystalline test
 Identification of blood can be made more specific if this test is applied or performed.
A. Teicmann Haemin test
B Acetone-Haemin test
C. Haemochromogen crystal test or Takayama test

MICROCHEMICAL TEST AND MICROCRYSTALLINE TEST

 Principle of the Test Test depends on the addition of the specific chemicals to the blood so that
characteristics crystals with hemoglobin will be formed

A. Teicmann Haemin test or Teicmann Test or Haemin Crystal test

Positive Result dark brown rhombic crystals arranged singly or in cluster
B. Acetone-Haemin test
Positive result: small dark, diachromic circular crystals
C. Haemochromogen crystal test or Takayama test
A delicate test for the presence of hemoglobin.
Positive Result large rhombic crystals of a salmon pink color arranged in cluster.
3. Spectroscope test
• Most delicate and reliable test for the determination of the presence of blood in both old and recent stains
• Perform using an optical instrument called spectroscope

PRECIPITIN TEST - determines whether the stain is human or animal in origin, if not human, the specific
animal family from which it is originated.
PRECIPITIN TEST FOR BLOOD
• Standard test used to determine whether the stain blood is human or animal in origin
• Very sensitive and requires only a small amount
• Stain dried as long as 10-15 years still give positive result
• Limitations precipitin reacts not only with blood proteins but also with other body proteins in saliva, semen,,
mucus and other body fluids.

BLOOD GROUPING TEST - determined the blood group if stain is human blood
BLOOD GROUPING OF FRESH BLOOD
If the stain is human blood, did it come from the victim, the accused or from other persons. The origin of
the blood or blood stain can be determined by the identification of the blood groups to which it belongs? This
identification is carried out on both fresh blood and blood stains
Human blood of all races can be divided into definite groups.
A-B-O SYSTEM
• In blood grouping of fresh blood A-B-O system is used.
• It was Landsteiner in 1900, who discovered the four blood groups namely group A, B, O and AB
• He named the 4 groups based on the basis of the agglutinogen or antigen contents of the RBC
• Agglutinogens or Antigens are characteristic chemical structures or principles that are founds on the surface of
the RBC which stimulates the production of agglutinins
• There are 2 agglutinogens classified as Agglutinogen A and agglutinogen B.
• On the other hand serum contains proteins or principles known as antibodies or agglutinins, which cause
agglutination or clumping of RBC
• They are anti-toxin substances within the body, which reacts when confronted with a specific antigens to
protect the system.
 WHAT ARE BLOOD TYPES?
There are 3 alleles or genes for blood type A, B, & O Since we have 2 genes, there are 6 possible combinations.

BLOOD TYPES
AA or AO = Type A
ВВ ог ВО = Туре В
OO = Type O
AB = Type AB
HOW COMMON IS YOUR BLOOD TYPE?
PERCENTAGE DISTRIBUTION OF BLOOD TYPE WORLD WIDE
TYPE DISTRIBUTION RATIOS
0+ 1 person in 3 38 4% 46.1%
0- 1 person in 15 7.7% 46.1%
A+ 1 person in 3 32.3% 38.8%
A- 1 person in 16 6.5% 38.8%
B+ 1 person in 12 9. 4% 11 1%
B- 1 person in 67 1.7% 11 1%
AB+ 1 person in 29 3.2% 3.9
AB- 1 person in 167 0.7% 3.9

BLOOD TRANSFUSIONS
A blood transfusion is a procedure in which blood is given to a patient through an intravenous (IV) line
in one of the blood vessels. Blood transfusions are done to replace blood lost during surgery or a serious injury.
A transfusion also may be done if a person's body can't make blood properly because of an illness.

WHO CAN GIVE YOU BLOOD?

 People with TYPE O blood are called Universal Doners, because they can give blood to any blood type
 People with TYPE AB blood are called Universal Recipients, because they can receive any blood type
Rh Can receive + or
Rb Can only receive
Rh FACTORS
• Scientists sometimes study Rhesus monkeys to learn more about the human anatomy because there are certain
similarities between the two species. While studying Rhesus monkeys, a certain blood protein was discovered.
This protein is also present in the blood of some people. Other people, however, do not have the protein.
• The presence of the protein, or lack of it, is referred to as the Rh (for Rhesus) factor.
• If your blood does contain the protein, your blood is said to be Rh positive (Rh+). If your blood does not
contain the protein, your blood is said to be Rh negative (Rh-).
TEN DIFFERENT MATINGS POSSIBLE BETWEEN THE 4 BLOOD GROUPS.
PARENTS POSSIBLE TYPE OF NOT POSSIBLE
CHILDREN
OxO O A, B, AB
AxO A, O B, AB
AxA A, O B, AB
BxO B, O A, AB
BxB B, O A, AB
AxB O, A, B, AB NONE
AB x O A, B O, AB
AB x A A, B, AB O
AB x B A, B, AB O
AB x AB A, B, AB O