FORENSIC

SEROLOGY

dr. Henky, Sp.F, M.BEth.

Department of Forensic Medicine

Faculty of Medicine Udayana University

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Daftar Ketrampilan Klinis (SKDI

2012)
Kedokteran Forensik
&
N
Keterampilan
o
Medikolegal

Tingkat
Keterampilan

Prosedur medikolegal

4A

Pembuatan Visum et Repertum

4A

Pembuatan surat keterangan medis

4A

Penerbitan sertifikat kematian

4A

Pemeriksaan selaput dara

Pemeriksaan anus

4A

Deskripsi luka

4A

Pemeriksaan derajat luka

4A

Pemeriksaan luar korban mati

4A

10 Pengambilan sampel vaginal swab, buccal

swab,

4A

darah, urine, dan isi lambung

11 Pemeriksaan bercak darah, cairan mani, dan
sperma

Locards Exchange Principle

Locards Exchange Principle
Every Contact Leaves a Trace
Every Contact Leaves a Trace
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Biological Evidence

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Forensic Analysis of Blood

1.
2.
3.
4.
5.

Visual examination of evidence

Presumptive screening test (Is it blood?)
Confirmation test (Seriously, is it blood?)
Determine species origin (human blood?)
Identify the blood (whose blood is it?)
SerologyTest
Test
Serology
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Species Origin (1)

Most methods test for serum proteins
Serum proteins are found in all animals,
but are slightly different
Species ID methods based on antigenantibody interactions

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Species Origin (2)

Antigen = Serum Protein
Antibody = Produced when
foreign serum protein is
detected
Certain antibody will only
attach to one speciess
serum protein

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Methods
1.
2.
3.
4.

Ring Precipitin Test (Uhlenhut)

Ouchterlony Double Diffusion
Crossed-Over Electrophoresis
BM Test Colon Albumin (Sudiono S)

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Ring Precipitin Test (1)

Making Human Antiglobulin Serum

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Ring Precipitin Test (2)

Blood sample (dilute)
in top layer
Antiserum in heavy
bottom layer

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Ring Precipitin Test (2)

Blood sample (dilute)
in top layer
Precipitate means blood &
antiserum species match

Antiserum in heavy
bottom layer
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Ouchterlony Double Diffusion

Human blood

Human
antiserum
Not human
blood

Antiserum placed in center

Several bloodstains tested at one time

White line means antiserum and blood match

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Crossed-Over Electrophoresis
Antiserum

Blood Stain
Gel
Holes

Antiserum and Blood move toward each other

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Crossed-Over Electrophoresis
Antiserum

Blood Stain

Antiserum and Blood move toward each other

If line forms, antiserum and blood match

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BM Test Colon Albumin

Blood Stain
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Floating Sol

BM Test Colon Albumin

(+): Red / Violet

Blood Stain
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Floating Sol

BM Test Colon Albumin

(-): Yellow / Orange

Blood Stain
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Floating Sol

Identify The Blood

If a stain is blood, and it is human blood,
then whose is it?
1. Blood Group Markers
2. Protein / Enzyme Markers
3. DNA

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Blood Group Markers

Erythrocyte surface antigen
ABO, MNS, Rhesus, Duffy, Kell, Kidd, etc

Protein serum system

Gm, Gc, Haptoprotein, etc

Erythrocyte enzyme system

PGM, AK, ADA, PCE, EAP, GPT, 6-GPD, etc

Leucocytes antigen
HLA

Others
Secretor/non secretor, platelets antigen
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ABO Markers (1)

Antigens on RBC are glycoproteins and
are attached to the outside of the cell

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ABO Markers (2)

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ABO Markers (3)

Testing is similar to species test
An anti-A, anti-B, or anti-AB antiserum
(containing antibodies) is reacted with the
blood stain to detect blood cells
A, B, or AB blood cells are reacted with a
blood stain to detect antibodies
Tests can get complicated with absorbing and
releasing cells
Final step is usually testing for agglutination
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Direct Test

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Indirect Test
(Absorption Elution)

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Protein or Enzyme Markers (1)

Some protein or enzymes can be in different
forms (different shapes)
These differences can be detected by
separating the protein in a gel by
electrophoresis

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Protein or Enzyme Markers (2)

+

Charge makes proteins

move through gel
Different shapes move
at different rates
After several minutes,
their location will tell
what type they are.
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Ladder Type 1 Type 2 Type 3

Protein or Enzyme Markers (3)

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Another Uses of Forensic Serology

Paternity Cases:
The babies get accidently exchanged
Kidnapped baby
Man suspected as biologic father
Husband confuse about the baby
Disputed paternity
Mendel Law
Paternity exclusion
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ABO Mendelian Inheritance

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Case (1)
Phenotypes
Phenotypes
Father

Mother

AB

O
Child
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Case (2)
Phenotypes
Phenotypes
Father

Mother

AB

AB
Child
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Percentage of Blood Types

Blood Type

Antigen
(Blood Group)

Antibody

% Population

Anti-B

40

Anti-A

10

AB

A&B

None

Anti-A &
Anti-B

45

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Blood Group Markers

Probability (%)

RBC Antigen
MNS

32.1

Rhesus

28.0

Kidd

19.0

Duffy

18.0

ABO

17.6

Kell

3.3

Lutheran

3.3

Protein Serum
Gc

24.7

Hp

17.5

Glm

6.5

Km

6.0

RBC Enzyme
PGM

25.3

EAP

21.0

GPT

19.0

Glyoxalase

18.4

Esterase D

9.0

AK

4.5

ADA

4.5

HLA

94.0

Rape
Serological evidence is crucial
Forceful physical contact
Transfer of evidence:
Blood
Semen
Perspiration
Urine
Hair
Nail
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Semen
Consist of:
1. Sperm cells
2. Seminal Fluid Protein Rich Bodily Fluid
Serology Tests for Semen are:
a. Acid Phosphatase Test
b. Prostate Specific Antigen (p30)

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Acid Phosphatase Test

Adam & Wraxall Gel Electrophoresis:
1. Polyacrylamide Gel
2. Buffer pH 8,5
3. Substrate: 4-methylumbelliferyl phosphate
4. Result:
Seminal Acid Phosphatase ANODA
Vaginal Acid Phosphatase KATODA

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PSA Test by Electrophoresis

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PSA Rapid Test Device

Membrane Based Immunochromatography

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Results

Invalid Results

Research
Year

Researcher

Rapid Test

Sen Spe PPV NPV

Time

2004

Khaldi, et al

PSA Check-1

99.4

98

99.4

98

48 h

2007 Peonim, et al One-Step PSA

85

85

89

79

2009

Hobbs, et al

ABA-Card

100

96

2009

Laux, et al

Seratec

7296 h

2013 Peonim, et al InTec Products 80.4 92.3 77.6 93.5

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If Semen, Whom Does it Belong to?

Blood Group Type
Secretor : ABO, Lewis Blood Typing
Methods: Absorption Inhibition

DNA : Y-STR
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Genetic Markers in Blood / Semen

ABO blood typing and protein analysis
may help eliminate a suspect (exclusion)
Since there are only a small number of
types (ABO = 4 types), a match does not
mean the stain definitely came from a
certain person
DNA testing is very specific, thus it can
identify a person, and is becoming just as
easy as the above tests
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THANK YOU