The Consequences of Using Different Storage Temperature of 5°C
and 10°C in The Identification of Blood Group ABO from Dental
Pulp Samples
Riki Kristanto
1
, Yanuar Kristanto
2
, Slamet Soetanto
3
1
Dentist, Master of Forensics Student, Universitas Airlangga, Surabaya, Indonesia.
2
Dentist, Master of Immunology Student, Universitas Airlangga, Surabaya, Indonesia.
3
Conservative Dentistry Specialist and Staff Lecture in Faculty of Dentistry, Univeristas Airlangga, Surabaya, Indonesia.
Keywords: Forensic Dentistry, ABO blood Group, Dental Pulp, Absorption-Elution, and Temperature.
Abstract: In forensic dentistry, the examination of the blood group ABO from dental pulp samples can be performed
on secretors, which relate to people who have antigens and antibodies in their body tissue constructions.
However, this case has several factors that can influence the results of the examination of the blood group
from dental pulps which only lasts 180 days. The objective is to identify the result differences in storage
treatment between the temperatures of 5°C and 10°C for the blood group analysis from the dental pulp
samples. This research analyzes blood by using the absorption-elution technique. The 18 dental pulp
samples taken from the ABO blood group. Each blood group consists of 6 samples. The research was
performed during the period of April-July 2012. The results indicate that there are similarities between the
blood pulp samples stored at 5°C and 10°C with the origin blood types. This is confirmed by using the one-
way ANOVA test, with p = .884. The examination of the blood group using the dental pulp in a storage
temperature of 5°C indicates an accurate result. However, the accuracy decreases if the storage temperature
is 10°C.
1 INTRODUCTION
In the investigation of a criminal act, one must
provide scientific physical evidence (Wirasuta,
2010). This evidence helps the investigator to
identify the perpetrator. Some techniques could be
applied, from simple techniques such as blood type
analysis, fingerprint analysis and dental
identification to a more sophisticated technique such
as DNA (deoxyribonucleic acid) analysis (Lukman,
2016).
In forensic dentistry, the blood type of an
individual person’s dental becomes a personal
identification. Theoretically, the blood type comes
from the antigen on the surface of a red blood cell.
This antigen can interact with a specific antibody
(Ballad, Davis, 2011). The utilization of the blood
type in medico-legal examination is the primary
method to identify individuals (Shetty, 2010).
The sample of a blood type analysis could be
obtained from the dental pulp. There are several
steps involved, such as sample extraction,
preservation and sample storage. To obtain an
optimum result, the analysis should be conducted in
a short storage time at 5°C (Lukman, 2006).
According to Alcemo (1985), the storage of all
specimens could be stored at a low temperature
between 4°-10°C. However, if the dental pulp
storage duration is too long, it will cause the
denaturation of the sample by a chemical and
microbiological process which will influence the
sample’s integrity (Shetty, 2010).
Based on that background, the aim of this
research is to discover the different outcome on the
conditions of the ABO blood type from the dental
pulp samples when stored in temperatures between
5° and 10°C.
2 MATERIALS AND METHODS
Permanent dental pulp was used as a sample in the
experiment. This dental pulp originated from dental
extraction in Bhayangkara Hospital in Kediri City.
Kristanto, R., Kristanto, Y. and Soetanto, S.
The Consequences of Using Different Storage Temperature of 5Â
ˇ
rC and 10Â
ˇ
rC in The Identification of Blood Group ABO from Dental Pulp Samples.
DOI: 10.5220/0007543804030406
In Proceedings of the 2nd International Conference Postgraduate School (ICPS 2018), pages 403-406
ISBN: 978-989-758-348-3
Copyright
c
2018 by SCITEPRESS – Science and Technology Publications, Lda. All rights reserved
403
The research was conducted in April - July 2012.
The total respondents were 18 individuals. The
respondents were divided into 3 groups of 6 based
on their blood type: A, B, and O blood. This study’s
research used the Absorption-Elution method. For
the data processing and analysis, One-way ANOVA
was used. Moreover, Post Hoc LSD was applied to
find the difference between the groups.
Laboratory analysis (Absorption-Elution
method) is conducted as follows (Lukman, 2006):
The tooth that still have pulp tissue was taken,
then was ground on iron mortar until it became
powder. The tooth powder was placed into 3 test
tubes. 3 different antisera (α to tube I, β to tube II, γ
to tube III). All tubes were stored in a refrigerator at
a certain, temperature for 24 hours (overnight), then
the reaction was rinsed with a saline solution 7 times
for each test tube.
From each test tube the saline solution was
removed, but not the precipitate, and put into the 3
test tubes. 2 drops of aquadest were added. The test
tubes were heated up to 56°C for 12 minutes using a
microwave, then the test tubes were taken out from
the microwave.
The indicator cells A, B, and O were inserted
into each test tube with a concentration of 3 - 5%
each, then, the test tubes were centrifuged until
agglutination occurred, in the end, the tubes were
observed for when the agglutination occurred, these
steps were conducted in the storage condition of 5°C
and 10°C.
3 RESULT
The research was conducted in April - July 2012 in
the Hematology Laboratory of Baptist Hospital in
Kediri City. The total respondents were 18
individuals. The respondents were divided into 3
groups of 6 based on their blood type, A, B and O.
The result from the two storage conditions (5°C
and 10°C) is summarized in Table 1.
Table 1. Blood type examination result
N
o
Original Blood
type Respondent
Blood type from Dental
pulp Examination
Temp. 5°C
Temp.
10°C
1
A
A
A
2
A
A
B
3
A
A
A
4
A
A
A
5
A
A
-
A
A
A
B
B
B
B
B
B
B
B
B
B
B
B
B
B
-
B
B
B
O
O
O
O
O
O
O
O
O
O
O
-
O
O
O
O
O
-
All data including the original blood type of the
respondents (research control) and the blood type
from the dental pulp examination in both storage
temperatures (5°C and 10°C) were processed by the
parametrically statistic method (One-way ANOVA).
This processing method is used to evaluate if the
results are significant enough between the two
storage conditions and research control.
A statistical data analysis was performed using
SPSS software. In the Normality Shapiro-Wilk Test,
the significant value showed .097, .089, and .055 so
that p value > 0.05. It means all data was distributed
normally. The value in homogeneity of the variance
test showed .051 so that, p-value > 0.05. It means all
treatments have the same variation in this blood type
examination. These results met the requirements to
perform a One-way ANOVA test.
The result of the One-way ANOVA test showed
the mean square between the groups were .020 and
.160 and the p-value of .884. This value is more than
0.05. Therefore, it is concluded that the blood test
experiment results are significant enough among all
three of the treatments. However the mean result
was different between storage in 5°C and 10°C.
4 DISCUSSION
Figure 1. The Result of ABO blood type from dental pulp.
ICPS 2018 - 2nd International Conference Postgraduate School
404
In this research it is known that dental pulp could
become the material used in ABO blood type
determination by using the absorption-elution
technique at a storage temperature of 5°C and 10°C.
These findings align with previous research
conducted by Inamdar (2011) which showed that the
ABO blood type identification was performed by
using the same technique. This technique has been
applied in Forensic Science for determining the
ABO blood type. Moreover, this technique must use
a specific temperature.
According to Alfonsius and the research team
from Police Medical Laboratory in 1992, ABO
blood type examination from a dental pulp sample
could be conducted by using the absorption-elution
technique which is should be stored at 5°C (Lukman,
2006).
Many research results align with the theory that
states dental pulp could be utilized for ABO blood
type examination only for 180 days since the date of
death. (Shetty, 2010; Ballal and David, 2011).
Based on the blood type examination data, it was
known that the dental pulp sample examination after
storing it at 5°C resulted in the same result as the
original blood type. It was caused by the ability of
the temperature to reduce microbiological growth
and maintain optimum specimen integrity. In dental
pulp there are some types of matrix that produce
plasma cell specification such as glycosaminoglycan
and fibronectin (Goldberg and Smith, 2004). These
substances will not denature so that the blood type
analysis and the original blood type sample results
end up being the same.
Different findings were encountered when the
dental pulp sample was stored at 10°C. This
treatment resulted in different blood types from the
original blood types of the sample. It is because the
growth of decaying organisms becomes faster at
10°C (Biogenex Laboratories, 2006). It will denature
the dental pulp specimen through a chemical and
microbiological process then the specimen integrity
will be altered leading to alteration of the
specimen’s integrity. These findings align with
previous research, conducted by Inamdar (2011). It
stated that the inaccuracy of blood type examination
was caused by an aerobic microorganism such as
gram-negative bacteria (E.coli and S. Maracessens).
These bacteria may alter the antigen determination if
the dental pulp sample becomes a B antigen which
can influence the results. However, according to
Ballal and David (2011), the negative results in the
blood type examination was caused by an aerobic
gram-negative bacteria specimen contamination.
The One-way ANOVA test shows the p-value of
.884 which is more than 0.05. It means there is a
similarity between the original blood type of the
sample and the experimental blood type of the dental
pulp sample which was stored at 5°C and 10°C.
5 CONCLUSIONS
From the research results of the ABO blood type
examination using a dental pulp sample in the
Hematology Laboratory of Baptist Hospital in
Kediri City, it is concluded that a blood type can still
be determined by using a dental pulp sample stored
at either 5°C or 10°C. However, storing a dental pulp
sample in the temperature of 5°C gives a result
closer to the original blood type compared to storing
the dental pulp sample at the temperature of 10°C.
The result of storing the dental pulp sample at
10°C showed 4 undetectable samples, because the
specimen can not be maintained well at that
temperature. The effect of this is the denaturation of
the dental pulp specimen through a chemical and
microbiological process which at that point the
specimen’s integrity is altered. This resulted in the
agglutination not occurring.
REFERENCES
Alcamo, 1985, Fundamentals of Microbiology, Addison-
Wesley Publishing Company, Sydney, pp. 234-
236.
Ballal, S., and David, MP., 2011, Determination of ABO
Blood Grouping From Dentine and Pulp, Pakistan
Oral and Dental Journal, 31(1):3-6.
BioGenes Laboratories Inc, 2006, Antigen Retrieval,
accessed on
http://www.biogenex.com/customercare/faq_antig
en.asp.
Goldberg, M., and Smith, A.J., 2004, Cells and
Extracellular Matrices of Dentin and Pulp: A
Biological Basis for Repair and Tissue
Engineering, International & American
Association for Dental Research, Volume: 15
issue: 1, page(s): 13-27.
Inamadar, P., 2011, Teeth-Hidden Treasure of Blood
Group, International Journal of Advanced
Research on Oral Sciences, 1(1):5-8.
Lukman, D., 2006, Buku Ajar Ilmu Kedokteran Gigi
Forensik, Jilid 2, Cv. Sagung Seto, Jakarta, pp. 1-
2, 77-80.
Shetty, M., 2010, ABO Blood Grouping from Tooth
Material, Journal Indian Acad Forensic Med.,
32(4):336-338.
The Consequences of Using Different Storage Temperature of 5Â
ˇ
rC and 10Â
ˇ
rC in The Identification of Blood Group ABO from Dental
Pulp Samples
405
Wirasuta, I., 2010, Pengatar Menuju Ilmu Forensik, Bahan
Kuliah Forensik, accessed on
www.scribd.com/doc/27116840/Pengantar-
Menuju-Ilmu-Forensik,
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