JCDR 10 ZC01
JCDR 10 ZC01
JCDR 10 ZC01
7161
Original Article
ABSTRACT Materials and Methods: Blood and saliva samples were collected
Introduction: The ABO blood group system was the significant from 80 individuals comprising 20 individuals in each blood group.
element for forensic serological examination of blood and body The absorption inhibition method was used to determine the blood
fluids in the past before the wide adaptation of DNA typing. A group antigens in the saliva and then the results were correlated
significant proportion of individuals (80%) are secretors, meaning with the blood group of the collected blood sample. The compiled
that antigens present in the blood are also found in other body data was statistically analysed using chi-square test.
fluids such as saliva. Absorption inhibition is one such method that Results: Blood groups A & O revealed 100% secretor status for
works by reducing strength of an antiserum based on type and both males and females. While blood groups B and AB revealed
amount of antigen present in the stains. 95% secretor status.
Aim: To check the efficacy of identifying the blood group antigens Conclusion: Secretor status evaluation of the ABO blood group
in saliva and to know the secretor status using absorption inhibition antigen in saliva using absorption inhibition method can be a useful
method among southern Rajasthan population. tool in forensic examination.
Keywords: Body fluid, Blood stain, Forensic identification, Haemagglutination, Saliva analysis
vein of right arm, centrifuged for five minutes to collect pure form Blood groups Total number of
of indicator erythrocytes and at the same time 4 -5 ml of whole study subjects Males Females
unstimulated saliva was collected in a test tube by bending their A 20 15 5
heads for five minutes and directly collecting the saliva into the B 20 15 5
container. Blood grouping was done for the collected blood by ABO
AB 20 14 6
blood typing and estimation of salivary blood group antigens was
O 20 15 5
done by standard Absorption Inhibition Method.
Total 80 59 21
Procedure for Absorption Inhibition Method [Table/Fig-1]: Gender distribution of study subjects among each blood Group.
Test tubes with saliva were placed in boiling water bath for 10
minutes and allowed to cool. Cooled test tubes were centrifuged
for 10 minutes at 3000 rpm. After discarding the supernatant, clear
saliva was collected using pipette. For control test tube we added
one drop of saline in each.
Four test tubes were taken with 2 each labeled as TEST and
CONTROL. Control tubes were taken to ensure that the antisera were
not diluted beyond its capacity to agglutination. Stock agglutinating
reagent was carefully adjusted to 1:8 titers and one drop of diluted
antisera was added to each tube respectively. To every TEST tube,
one drop of clear saliva and to CONTROL tube one drop of saline
were added, later mixed, and finally incubated at room temperature
for minimum of 10 minutes. Then one drop of appropriate indicator
erythrocytes was added to each tube, mixed and incubated at room
temperature for 10 minutes. For saline reaction in the CONTROL,
tubes were centrifuged for 10 minutes [6-9].
Reaction for agglutination was recorded. Negative reaction test [Table/Fig-2]: Percentage distribution of the secretors and non secretors among
were re-evaluated using same procedure, and if negative again it different blood groups.
was considered as negative.
All control samples showed clumping as there was no antigen Sample
Author name used Method used Result
present. The test group was considered as positive if agglutination
Pawan saliva Absorption inhibition Out of 200 samples 83%
was not seen, indicating that antigen-antibody reaction had taken
motghare et method were secretors. Among
place between saliva and antisera, and there was no antibody left al., (2011) [9] females 83.8% showed
for RBCs to react, indicating the presence of blood group and vice secretor antigen and
81% in males.
versa was applied for negative test group samples.
Guriender kaur saliva and Inhibition and 87.5% were secretors in
et al., (1988) sweat stain Elusion method. saliva. 21.94% in sweat
RESULTs [12] with inhibition and 95.8%
Out of 80 subjects 59 were males and 21 females [Table/Fig-1]. with elution method.
A slightly higher percentage of secretor status was observed in M.H Graves vaginal Inhibition method Out of 25 samples 12
females (100%) than in males (90%). Groups A and O revealed (1978) [13] sample and Elusion method were of O group with 9
(75%) secretors and 13
100% secretor status for both males and females, while groups B were of A group with 8
and AB revealed 95% secretor status [Table/Fig-2]. (61.5%) secretors.
Robert Thaler saliva Haemagglutination out of 40 samples 31
DISCUSSION et al (1976) [8] Inhibition tests were (77.5%) were
secretors
After the advent of blood group system by Karl Landsteiner in
1900, the ABO blood group and Rh system distributions have Saboor M et saliva Haemagglutination 64.4% were secretors
al., (2014) [14] Inhibition tests while 35.6% were non-
shown marked variation around the world and also some variation secretors. Blood group B
in different areas within the same country. It has been reported by has the highest secretor
(79.5%)
various studies that the O blood type being common in American,
frequency while Blood
Canadian and Saudi population, the B type in Chinese and Indian group AB has the lowest
individuals, and the A type in Eskimos [8,10,11]. (45.5%)
The source of blood group antigens is the water soluble [Table/Fig-3]: Studies on secretary status of blood group antigens In various body
fluids using different methods.
substances that are present in most body fluids and organs of a
secretor. Yamakami discovered the presence of A and B antigens
in saliva [8]. This gave way to note in 1930 by Lehrs and Putko that from 25 families. The saliva was tested using absorption-inhibition
secretors or non secretors existed for these antigens as two distinct method. They found 83% secretors in their study subjects of which
groups [8]. With the concept once a blood group is established 83.8% were female secretors and 81% were male secretors [Table/
in an individual it remains unchanged throughout his life, the use Fig-3].
of blood group substances is based in medico legal examination. Kaur G et al., conducted a study on both saliva and sweat stains
Blood group would be determined with 100% accuracy from the and observed that 87.5% were secretor in saliva out of 72 samples
saliva of those subjects who were secretors of antigens in their tested [12] [Table/Fig-3]. In 1978 MH Graves studied 25 vaginal
saliva [4]. samples among which 17 (68%) were secretors of blood group
Finding of 99% secretor in this study is higher than the studies antigens [13] [Table/Fig-3]. Out of which 75% were of group O
conducted by Motghare P et al., who conducted a study in and 61.5% in group A [Table/Fig-3]. These two studies compared
Maharashtra population to determine blood group from saliva and Absorption-Inhibition and Absorption-Elution Methods in the
to know the secretary status [9]. Here they studied 200 subjects detection of ABO (H) antigens in vaginal samples, submitted in
(101 males and 99 females), 100 randomly selected and 100 taken sexual offense cases and stated that the Absorption-inhibition is a
well-established method for the identification of ABO (H) antigens class, and history can be formulated based on mere examination of
[14]. the teeth. At last correlating these evidences with those from forensic
Thaler R et al., studied saliva samples to determine if salivary blood investigators the identity possibilities can be narrowed down.
group substances are related to periodontal disease [8]. Forty
patients were studied, among which 31 (77.5%) were secretors CONCLUSION
for blood group antigens and in their study no correlation of the In the present study 100% secretor status was observed with blood
periodontal indices to blood group secretor status was established groups A and O for both males and females. The identification of
[Table/Fig-3]. blood group antigens in various body fluids like saliva chiefly, would
be a useful tool in forensic examination. Although recent techniques
In the most recent study done in 2014 by Saboor M et al., to
have increased the reliability of the determination of blood group
evaluate the ABH blood group among 101 healthy adult students
substances, adaption of the more sensitive assays may increase
(76 male, 25 female ranging in age from 15 to 40 years), to know the
the utilization of these samples. Of particular value would be the
secretors and non-secretors in people of Karachi, Pakistan using
development of simpler methods for blood group substances in
haemagglutination inhibition method of saliva [14]. They concluded
saliva since the antigens present in the blood are also found in other
that frequency of ABH secretor is high (64.4%). Blood group B has
body fluids as saliva.
the highest secretor (79.5%) frequency while blood group AB has
the lowest (45.5%). This result was correlated with ours.
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PARTICULARS OF CONTRIBUTORS:
1. Professor and Head of Department, Department of Oral & Maxillofacial Pathology, Pacific Dental College, Udhaipur, India.
2. Senior Lecturer, Department of Department of Oral & Maxillofacial Pathology, Institute of Dental Sceinces, Jammu, India.
3. Post Graduate Student, Department of Oral & Maxillofacial Pathology, Pacific Dental College, Udhaipur, India.
4. Reader, Department of Oral & Maxillofacial Pathology, Rama Dental College Hospital and Rescearch Center, Kanpur, Utter Predesh, India.
NAME, ADDRESS, E-MAIL ID OF THE CORRESPONDING AUTHOR:
Dr. Rashmi Metgud, Date of Submission: Oct 07, 2014
Professor and Head of Department, Department of Oral & Maxillofacial Pathology, Pacific Dental College, Udhaipur, India. Date of Peer Review: Jan 01, 2015
E-mail: [email protected] Date of Acceptance: Sep 21, 2015
Financial OR OTHER COMPETING INTERESTS: None. Date of Publishing: Feb 01, 2016