MEDICOLEGAL ASPECTS OF

BLOOD AND OTHER STAINS

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

 Different stains having
Medicolegal importance
 Blood
 Seminal fluid
 Vaginal discharge
 Saliva
 Faecal stain
 Urine

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

Blood stains
-Main cellular content RBC – Haemoglobin which
possess peroxidase activity
-Haemoglobin and other proteins of blood has
antigenic properties – produces antibodies when
injected into animals

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

 Medicolegal importance
 May establish a link between offence, offended
and offender
 May establish link between offence and offensive
agent
 Fictitious charges by presenting an animal stain
as human
 In poisoning, poison chemically detected
 Time since death may be estimated by chemical
and enzymatic study

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

 Examination
 Whether the stain is actually a blood stain
• Physical
• Chemical
• Microscopic
• Spectroscopic
• Micro chemical

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

 Whether the blood stain is of human or other than
human
• Precipitin test
 If human, which blood group
 Sex, source
 Indication as to cause of death
 Whether belong to victim or accused
 Whether bleeding antemortem or postmortem
 Any disease process

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

 Substances resembling blood stain
 Vegetable stains
- ammonia greenish yellow color
- Bleached by chlorine water
- M/E vegetable cells and fibers
 Rust stains
- Does not fall off in scale on being heated
- Positive reaction for iron
- Dissolved in dil. Hcl

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

 Mineral stains
 Synthetic dye stains
- Nitric acid yellow
strong alkali
original color
 Other stains

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

 Chemical examination
 Highly sensitive
 Positive reaction even with faint traces of blood
 Principal factor – peroxidase activity liberating
nascent oxygen which leads to colour change

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

1. BENZIDINE TEST

stain extract ┼ benzidine ┼ 10 vol

in hydogen peroxide
glacial acetic acid

Blue colour

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

2. KASTLE MAYER TEST
- Phenolphthalein in alkaline medium
- Pink colour
3. ORTHOTOLIDINE TEST
- (4% orthotolidine in ethyl alcohol)+ glacial acetic
acid + H2O2
- Green colour
4. LEUCHOMALACHITE GREEN TEST
- Bluish green colour
5. LUMINAL TEST

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

Confirmatory tests for blood

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

 Microscopic test
 RBC’s- confirmatory
 Species
 Sex
 Disease

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

 Microchemical test
 HAEMIN CRYSTAL TEST/ TEICHMAN’S
TEST
- Sod. Chloride + strain extract + glacial acetic acid
- Yellowish brown rhombic crystals of haemin
 HAEMOCHROMOGEN CRYSTAL/
TAKAYAMA TEST
- takayama reagent + stain extract
- Salmon pink hemochromogen crystals

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

 Spectroscopic examination
 Detects blood where RBC structure is lost and
hemoglobin has undergone chemical composition

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

 Species specification test
 PRECIPITIN TEST
- When human serum is injected in an
experimental animal, antibodies develop.
- When any human serum is brought in contact
with this animal serum, the antibodies and
protein will react and a visible precipitate will
develop

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

Identifying sex from blood stain
- Barr bodies in > 3% of polymorphs
- Y chromosome is fluorescent to quinacrine

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

Individuality by blood stain
Blood grouping
• red cell antigen
•White cell antigen
•Se protein polymorphism
•Red cell enzyme polymorphism

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

 Red cell antigen
 ABO system
 Rh system
 MNSs group system
 Lutheran
 Duffy
 Lewis
 Kidd

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

 secretors  Agglutinogens in water soluble form
 Secrete ABO antigens into body fluids and
Group specific substances

secretions
Non secretors

 Agglutinogens in lipoidal form

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

 White cell antigen
 Human Leucocyte Antigen (HLA)- the MHC in
humans
 Present on Chromosome 6 and encode cell
surface antigen presenting proteins
 4 types – HLA-A, HLA-B, HLA-C, HLA-D

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

 Serum protein polymorphism
 Occurrence in the same population of two or
more alleles at one locus, each with appreciable
frequency, where the minimum frequency is
taken as 1%
 Se. proteins subject to genetic variation are
• Se. haptoglobins
• Se lipoproteins
• Abnormal haemoglobins

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU

 Red cell enzyme polymorphism
 > 250 proteins & enzymes found in red cell
 Established value in paternity testing
• Phopho glucomutase
• Adenylate kinase

Dr. Shiuli, Forensic Medicine & Toxicology, KGMU