abo blood typing from forensic materials - merits and demerits of detection methods utilized in our...
TRANSCRIPT
![Page 1: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/1.jpg)
ABO Blood Typing from Forensic M - aterials Merits and demerits of de tection methods utilized in our lab
oratories, and biological significan ce of the antigens
By: Wiraya NakkeamkeaoAdviser: Dr.Sirilak Teeraputon
![Page 2: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/2.jpg)
Researcher
Katsuji Nishi, Steve n Rand,
Tokiko Nakagawa, Aiko Yamamoto,
Shigeru Yamasaki,Yoshio
Yamamoto, Aki Kobayashi,
Masateru Kane, Aki ra Morimoto,
Hedwege Spal thoff a ndBeate
Annuss
Journal of Forensic Medicine and Toxicology ,2005
![Page 3: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/3.jpg)
Introduction
- 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 method
![Page 4: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/4.jpg)
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/antigens.jpg
![Page 5: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/5.jpg)
ReviewABO Blood Groups
Blood Type
Genotyp
e
RBC Antige
n Presen
t
Serum Antibo
dy
A AA, Ai
A, H Anti-B
B BB, Bi
B, H Anti-A
AB AB A, B, H NoneO ii H Anti-A,
Anti-B
![Page 6: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/6.jpg)
ReviewSubgroups A
Group
Anti-A
Anti-B
Anti-AB
Anti-A1
A1 + - + +
A2 + - + -Anti-A1 = Lectin (Dolichos
biflorus)
![Page 7: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/7.jpg)
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
![Page 8: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/8.jpg)
ABO blood grouping method
Hemagglutinatio
n Absorption -
Elution Histochemical
techniques ABO
genotyping method
MethodMethod
![Page 9: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/9.jpg)
Hemagglutination method
A reaction outside the body between like antigen and antibody
- Cell grouping
- Serum grouping
“Hemagglutination” mean
![Page 10: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/10.jpg)
Hemagglutination methodPrinciple
![Page 11: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/11.jpg)
Hemagglutination method
![Page 12: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/12.jpg)
Hemagglutination method ABO grouping can be easily pe
rformed from corpses when f resh blood can be obtained
S torage periods of blood or in the postmortem time of corpse
s increases
1. C hanges in the agglutinabili ty 2. Deformability 3 . A ntibody binding capacity
4 . O smotic fragility 5 . M orphology
6 . ATPcontent
![Page 13: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/13.jpg)
Hemagglutination method
Developed Hemagglutination method - G lutalaldehyde or heat,an
- d storage in acidic dextrose (AC D) solution, blood bank conditio
n - B lood bank condition
and Incubation of the aged erythroc ytes in a saline solution containi
ng 5 mM adenine, 20 mM inosine and 5 mM glucose for 60 min at
370 C
![Page 14: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/14.jpg)
Hemagglutination method
Scanning electron microscopic observation of erythrocytes. Erythrocytes stored for 8 weeks in blood bank condition were incubated in the solution containing adenine and inosine. A; before incubation, B; after incubation.
http://www.geradts.com/anil/ij/vol_006_no_002/papers/paper001.html
![Page 15: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/15.jpg)
Hemagglutination method
Incubation period (min)
ATP content (mg/dl)
Agglutinability*
anti B Anti N Anti D
0 6.6 32 2 1
30 14.9 64 4 1
60 21.9 128 8 2
120 24.9 128 8 2
Table. Relationships between ATP content and agglutinability of erythrocytes stored 8 weeks
![Page 16: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/16.jpg)
Hemagglutination method 1 1. ml of blood sample was gentl
ii i i i ii i i i iii5 1 4 0 %iiiiiii iiiiiiii
2 . 6 2 5 %adjusted to pH with ci i i i ii i i iiiiiiii, 5 7 %
sesolution(pH6) was added. 3 . The sample was left standing for
5min,then washed three times with phy siologicalserum(i nact i vat ed gr oup AB human ser um) 4 . U 2sed to prepare a % red bloo
d cel l suspensi on
MichailowMichailow 's method 's method
It is often seen that the hemolys is of aged erythrocytes
![Page 17: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/17.jpg)
Absorption El ution methodPrinciple
![Page 18: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/18.jpg)
Absorption Elu tion method ABO typing of er ABO typing of er
ythrocytesythrocytes- Blood Stained * O ften encountered discrepant result
s when d etermining the blood group- M aking autopsies of bodies who
have died due to fire * G lycolipids and sialoglycoprot
eins
![Page 19: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/19.jpg)
Absorption Elu tion method
ABO typing fro ABO typing fro m body fluid m body fluidTable. Types of organs and tissues
expressing the ABH antigen
![Page 20: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/20.jpg)
Absorption Elu tion method ABO typing fro ABO typing fro
m body fluid m body fluidTable. Types of organs and tissues expressing the ABH antigen
![Page 21: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/21.jpg)
Absorption Elu tion method ABO typing fro ABO typing fro
m body fluid m body fluid- ABH and related antigens are
expressed in the secretory organs of many species of animals
- The ABH and their related anti gens were expressed in the sali
vary glands, kidneys and reprod uctive organs of animals as well as in those from humans
- The emphasize that the specie s identification prior to ABO blo
od grouping is indispensable
![Page 22: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/22.jpg)
Absorption Elu tion method ABO typing fro ABO typing fro
m other tissue m other tissue Skin Nail Hair Bone
www.cc.gatech.edu & www.emedicinehealth.com
![Page 23: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/23.jpg)
Histochemic al method
http://www.histochem.org/
- Material of body
- Staining ; Fluores
cent Dyes
- Microscopic
![Page 24: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/24.jpg)
Histochemic al method ABO Blood grouping from mate ABO Blood grouping from mate
rials obtained from humans rials obtained from humans
The antigens are easily and cert ainly detected by means of
immunohistochemical techniqu e using
* A ntibodies against ABH antigen s
* L enctins specific to blood group A, B, or O
It is a great merit that we can m icroscopically observe the
distribution of antigens and re action sites in the tissue
![Page 25: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/25.jpg)
ABO blood group incom ABO blood group incom patible cases patible cases
Histochemi cal method
ABO group determination in the blood group in compatible case. The transfused A typed red bl
ood cells indicated by arrows were easily detec ted inside blood vessels of the kidney with anti
A antibody (photo A), and a large amount of O t yped red cells shown by arrow inside blood ves
sels and collecting tubules were stained by ant i H antibody (photo B).
http://www.geradts.com/anil/ij/vol_006_no_002/papers/paper001.html
![Page 26: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/26.jpg)
Histochemi cal method ABH antigens in h
-air by animmuno his tochemical methodu
-sing the Avidin Bioti- n Peroxidase techni
.
HaiHairr
Figure . ABO grouping from hair. A-antigen was detected only in the medulla of the blonde hair showing mosaic pattern distribution. The
positive and negative sits in the medulla are observed by turns.
http://www.geradts.com/anil/ij/vol_006_no_002/papers/paper001.html
![Page 27: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/27.jpg)
Histochemi cal method Typing of Blood Gro Typing of Blood Gro
up A Subgroup up A Subgroup
- M onoclonal anti H antibody usuall y can notbind to the secretory cells,
epithelium of Hassall's corpuscles or blood vessel of A1 individuals
- T hese cells of A2individuals react - ed strongly with H antibody A2 in
dividuals may therefore be most cle arly discriminated from A1 individu al s by the di ff erence i nthe reac
ti vi ty of anti bodi es agai nst Aa ndHanti gens i nthe ti ssue secti
ons
![Page 28: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/28.jpg)
Histochemica l methodTypingTyping of secre of secre tor status tor status
- C ompared the histochemi cal properties of mucous cells
of sublingual and submandibul ar glands from blood group A a - nd AB Japanese non secretors
-with those from German non secretors
- A - nti A antibody, HPA an d HAA lectins
- P roved the ethnical diffe
rence in the expression of bloo d group antigens
![Page 29: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/29.jpg)
Histochemi cal method ABO Blood grouping from ABO Blood grouping from
materials obtained from cri materials obtained from cri minal scene minal scene
- ABH antigens are expressed in the secretory organs of man
y species animals - D istribution of ABH antigen
s on organs and cells in each a nimal or human is revealed an
d the reaction sites on the tiss ue sections can be microscopic
ally
![Page 30: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/30.jpg)
Histochemi cal methodTable . Expression patterns of ABH antigens and lectin binding in red cells,
endothelial cells,and secretory cells of various animal species
![Page 31: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/31.jpg)
Histochemi cal methodTable . Expression patterns of ABH antigens and lectin binding in red cells,
endothelial cells, and secretory cells of various animal species
![Page 32: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/32.jpg)
Histochemic al 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 sho
wn 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_002/papers/paper001.html
![Page 33: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/33.jpg)
ABO genotyping
Method1. DNA extraction
2. Master mix preparing and PCR reaction
3. Amplification4. Gel electrophoresis5. Ethidium staining
6. Gel document
![Page 34: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/34.jpg)
ABO genotyping
http://wheat.pw.usda.gov/~lazo/docs/cotton/image/pcr.jpg
![Page 35: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/35.jpg)
ABO genotyping
- DNA extracted and fresh blood and formalin-fixed paraffin embedded
tissues. - Amplified by
PCR Figure. Electrophoretic pattern of the PCR produ
ct 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(Wist ar),8;Rat(Fischer), 9; Mastomys, 10;Mouse, and
11: Goldfish
![Page 36: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/36.jpg)
Discussion - ABH and related antigens may b
-e associated with the chemo recepto r system
- ABO polymorphism might appea
r in contact with the external enviro nment and play a role in the relation
ships of the species with pathogens, although the antigen might be invol
- ved in cell cell recognition in the em bryo in lower vertebrate species
![Page 37: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/37.jpg)
Conclusion
![Page 38: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/38.jpg)
Conclusion
![Page 39: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/39.jpg)
References1. ABO Blood สื�บค้�นข้�อมู�ลเมู��อวั�นที่�� 8 กรกฎาค้มู 2549 http://www.mie.utoronto.ca/labs/lcdlab/biopic/fig/1014. .jpg
2. ABO Blood สื�บค้�นข้�อมู�ลเมู��อวั�นที่�� 8 กรกฎาค้มู 2549 http://www.enh.org/uploadedimages/antigens.jpg3. ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens สื�บค้�นข้�อมู�ลเมู��อวั�นที่�� 15 มู�ถุ�นายน2549 http://www.geradts.com/anil/ij/vol_006_no_002/papers/paper001.html4 . ศาสืตราจารย�แพที่ย�หญิ�งที่�ศน�ยาน� จ�นที่นย��งยง , เวัชศาสืตร�ที่การธนาค้ารเล�อด , เมูษายน 2541 , พ�มูพ�ค้ร�)งที่�� 1 ,ธรรกมูลการพ�มูพ� ,กร�งเที่พ5. Forensic science สื�บค้�นข้�อมู�ลเมู��อวั�นที่�� 1 กรกฎาค้มู 2549
http://www.barascientific.com/bscnews/forum/Forensice/forensic.php
![Page 40: ABO Blood Typing from Forensic Materials - Merits and demerits of detection methods utilized in our laboratories, and biological significance of the antigens](https://reader035.vdocuments.us/reader035/viewer/2022062423/56649edd5503460f94bedda6/html5/thumbnails/40.jpg)
THE END