Transcript ABO Blood Typing from Forensic Materials

ABO Blood Typing from Forensic
Materials - Merits and demerits of
detection methods utilized in our
laboratories, and biological significance
of the antigens
By: Wiraya Nakkeamkeao
Adviser: Dr.Sirilak Tee
Katsuji Nishi, Steven
Rand,
Tokiko Nakagawa, Aiko
Yamamoto,
Shigeru Yamasaki,
Yoshio
Yamamoto, Aki
Kobayashi,
Masateru Kane, Akira
Morimoto,
Hedwege
Spalthoff
and
Journal of Forensic Medicine and
Toxicology ,2005 Beate
Researche
r
Introducti
on
- In forensic science and medicine
the ABO system has been
a major
focus
- A, B and O(H) antigens on
erythrocytes are also associated
with
other cells and tissues throughout the
body
- A, B and O(H) antigens are known
to be considerably
stable to the such
violent conditions as heating or drying
- The merits and demerits of the
ABO blood-grouping
Objective
- The merits and demerits of the ABO blood grouping methods
- Discuss the biological significance of the ABH and its related
antigens on basis of the data showing the distribution of these
antigens in the animal organs
http://www.enh.org/uploadedimages/an
Review
ABO Blood Groups
Blood
Type
Geno
RBC
Serum
type Antigen Antibo
Present
dy
A
AA, Ai
A, H
Anti-B
B
BB, Bi
B, H
Anti-A
AB
AB
A, B, H
None
O
ii
H
Anti-A,
Review
Subgroups A
Grou Anti- Anti- Anti- Antip
A
B
AB
A1
A1
+
-
+
+
A2
+
-
+
-
Anti-A1 = Lectin (Dolichos biflorus)
Forensic
material
"Forensic material"
means:
(a) Sample ; blood, semen or
(b) Hand prints, finger prints,
foot prints
or toe prints; or
(c) Taken from or of a
person’s body by a forensic
procedure
ABO blood
 Hemagglutination
grouping
method
Method
 Absorption
Elution
 Histochemical
techniques
 ABO genotyping
method
Hemagglutinatio
“Hemagglutination”
mean
n method
A reaction outside the body
between like antigen and
antibody
- Cell grouping
- Serum grouping
Hemagglutinatio
nPrinciple
method
Hemagglutinati
on method
Hemagglutinati
on method
 ABO grouping
can be easily
performed from corpses
when
fresh blood can be obtained
 Storage periods of blood or in the
postmortem
time
of
corpses
increases
1. Changes in the agglutinability
2. Deformability
3. Antibody binding capacity
Hemagglutinati
Developed
Hemagglutination
method
on
method
- Glutalaldehyde or heat,and
storage in acidic-dextrose (ACD)
solution, blood bank condition
- Blood bank condition and
Incubation of the aged erythrocytes in
a saline solution containing 5 mM
adenine, 20 mM inosine and 5 mM
glucose for 60 min at 370 C
Hemagglutinatio
n method
Scanning electron microscopic observation of erythro
in blood bank condition were incubated in the solution cont
incubation, B; after incubation.
http://www.geradts.com/anil/ij/vol_006_no
Hemagglutinatio
Table. Relationships between ATP content
and agglutinability
of
n method
erythrocytes stored 8 weeks
Incubation
period (min)
ATP content
(mg/dl)
0
Agglutinability*
anti B
Anti N
Anti D
6.6
32
2
1
30
14.9
64
4
1
60
21.9
128
8
2
120
24.9
128
8
2
Hemagglutinatio
1 ml
of method
blood
sample
was gently
It is 1.
often
seen
that the
hemolysis
n
Michailow's
shaken
for 5 min in 1ml of 40% saccharose
of aged erythrocytes
solution
method
2. adjusted to pH 6 with 25% citric
acid, and then 5 ml of 7% saccharose
solution (pH 6) was added.
3. The sample was left standing for 5
min,then washed three times with
physiological serum (inactivated group AB
human serum)
4. Used to prepare a 2% red blood cell
Absorption
Principle
Elution
method
Absorption
ABO
typing
of
Elution method
erythrocytes
- Blood Stained
* Often encountered
discrepant results when determining
the blood group
- Making autopsies of bodies who have
died due to fire
* Glycolipids
and sialoglycoproteins
Absorption
ABO typing from
method
Table.
Types
of organs and
tissues
bodyElution
fluid
expressing the ABH antigen
Absorption
ABO typing from
Elution
method
Table.
of organs and
tissues
bodyTypes
fluid
expressing the ABH antigen
Absorption
ABO typing from
Elution
method
body fluid
- ABH and related antigens are
expressed in the secretory organs of
many species of animals
- The ABH and their related antigens
were expressed in the salivary glands,
kidneys and reproductive organs of
animals as well as in those from
humans
Absorption
ABO Elution
typing from method
other tissue
 Skin
 Nail
 Hair
 Bone
www.cc.gatech.edu &
www.emedicinehealth.com
Histochemical
- Materialmethod
of body
- Staining ;
Fluorescent Dyes
- Microscopic
http://www.histochem
Histochemical
 The antigens are easily and certainly
ABO
Blood
grouping
from materials
method
detected
by means
of
immunohistochemical
obtained
from humans
technique
using
*Antibodies against ABH antigens
*Lenctins specific to blood group A,
B, or O
 It is a great merit that we can
microscopically observe the
distribution of antigens and reaction
sites in the tissue
Histochemical
ABO
blood
group
method
incompatible cases
ABO group determination in the blood group
incompatible case. The transfused A typed red blood
cells indicated by arrows were easily detected inside
blood vessels of the kidney with anti A antibody (photo
A), and a large amount of O typed red cells shown by
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Histochemica
ABH antigens in hair
Hair
l
method
by
animmuno-
histochemical
methodusing
the
Avidin-BiotinFigure
. ABO grouping
from hair. A-antigen wa
Peroxidase
technique.
medulla of the blonde hair showing mosaic pa
positive and negative sits in the medulla are ob
http://www.geradts.com/anil/ij/vol_006_no
Histochemical
- Monoclonal
antiBlood
H antibody
usually
Typing of
Group
can not bindmethod
to the secretory cells,
A Subgroup
epithelium of Hassall's corpuscles or
blood vessel of A1 individuals
- These cells of A2 individuals reacted
strongly with H antibody A2
individuals may therefore be most
clearly
discriminated
from
A1
individuals by the difference in the
reactivity of antibodies against A and
Histochemical
Compared
the
histochemical
Typing of secretor
properties method
of mucous cells of
status
sublingual
and
submandibular
glands from blood group A and AB
Japanese non-secretors with those
from German non-secretors
- Anti A-antibody, HPA and
HAA lectins
Proved
the
ethnical
difference in the expression of
Histochemical
ABO Blood method
grouping from
materials obtained from
criminal scene
- ABH antigens are expressed in
the secretory organs of many
species animals
- Distribution of ABH antigens on
organs and cells in each animal or
human is revealed and the reaction
Histochemical
Table . Expression patterns of ABH antigens and
method
endothelial cells,and secretory cells of va
Histochemical
Table . Expression patterns of ABH antigens an
method
endothelial cells, and secretory cells of
Histochemical
method
Figure. ABO grouping from mummified skin. Endothelial cells of blood vessel in the mummified skin
obtained from the leg (photo B) of a skeleton body (photo A) showed reactivity with corresponding antibody
as shown in photo C. The PCR product of DNA obtained from the mummified skin could not be amplified .
http://www.geradts.com/anil/ij/vol_006_no
ABO
Method
genotyping
1. DNA extraction
2. Master mix preparing and
PCR reaction
3. Amplification
4. Gel electrophoresis
5. Ethidium staining
6. Gel document
ABO
genotyping
http://wheat.pw.usda.gov/~lazo/docs/cotton/im
ABO
- DNA extracted
genotyping
and fresh blood
and formalin-fixed
paraffin
embedded tissues.
- Amplified by PCR
Figure. Electrophoretic pattern of the PCR product using
pairs of primers for ABO genotyping. M;174/Hae III,
1;Human, 2;Japanese Monkey, 3;pig, 4;Cat, 5;Rabbit,
6;Chinese Hamster, 7;Rat(Wistar),8;Rat(Fischer), 9;
Discussion
- ABH and related antigens may be
associated with the chemo-receptor system
- ABO polymorphism might appear in
contact with the external environment and
play a role in the relationships of the
species with pathogens, although the
antigen might be involved in cell-cell
recognition in the embryo in lower
vertebrate species
Conclusion
Conclusion
References
่ นที่ 8 กรกฎาคม 2549
1. ABO Blood สืบค ้นข ้อมูลเมือวั
http://www.mie.utoronto.ca/labs/lcdlab/biopic/fig/10.14.j
่ นที่ 8 กรกฎาคม 2549
2. ABO Blood สืบค ้นข ้อมูลเมือวั
http://www.enh.org/uploadedimages/antigens.jpg
3. ABO Blood Typing from Forensic Materials - Merits and deme
laboratories, and biological significance of the antigens สืบค ้น
http://www.geradts.com/anil/ij/vol_006_no_002/papers/pape
่
4. ศาสตราจารย ์แพทย ์หญิงทัศน์ยานี จันทนยิงยง
, เวชศาสตร ์ทการธนาคา
้ั ่ 1 ,ธรรกมลการพิมพ ์ ,กรุงเทพ
พิมพ ์ครงที
่ นที่ 1 กรกฎาคม 2549
5. Forensic science สืบค ้นข ้อมูลเมือวั
http://www.barascientific.com/bscnews/forum/Forensice/fore
THE
END