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ScienceDirect
Available online at www.sciencedirect.com
Procedia Manufacturing 51 (2020) 975–980
2351-9789 © 2020 The Authors. Published by Elsevier Ltd.
This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/licenses/by-nc-nd/4.0/)
Peer-review under responsibility of the scientific committee of the FAIM 2021.
10.1016/j.promfg.2020.10.137
10.1016/j.promfg.2020.10.137 2351-9789
© 2020 The Authors. Published by Elsevier Ltd.
This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/licenses/by-nc-nd/4.0/)
Peer-review under responsibility of the scientic committee of the FAIM 2021.
Available online at www.sciencedirect.com
ScienceDirect�
Procedia Manufacturing 00 (2021) 000–000
www.elsevier.com/locate/procedia
2351-9789 © 2020 The Authors. Published by Elsevier Ltd.
This is an open access article under the CC BY-NC-ND license https://creativecommons.org/licenses/by-nc-nd/4.0/)
Peer-review under responsibility of the scientific committee of the FAIM 2020.
30th International Conference on Flexible Automation and Intelligent Manufacturing (FAIM2021)
15-18 June 2021, Athens, Greece.
Development of Prototype Kit for Portable Drug Allergy Testing
Somkiat Tangjitsitcharoen1*, Jettanong Klaewsongkram2,
Angsumalin Senjuntichai1, Supranee Buranapraditkun2, Sutthinee Thongnop1
1Department of Industrial Engineering, Faculty of Engineering, Chulalongkorn University, Thailand
2Department of Medicine, Faculty of Medicine, Chulalongkorn University, Thailand
* Corresponding author. Tel.: +6-689-388-1688. E-mail address: somkiat.ta@eng.chula.ac.th
Abstract
The prototype kit for portable drug allergy testing is proposed and developed for easy-to-use in provincial hospitals where the standard equipment
for in vitro drug allergy diagnosis is still lacking. The Enzyme-Linked Immunosorbent Spot (ELISpot) technique is adopted for this purpose due
to its high sensitivity and specificity to detect cytokine-releasing cells, which is precise and suitable for non-instantaneously allergic tests. A
developed prototype for drug allergy testing has been considered in various aspects of the three main designs, comprising of the proposed testing
procedure kit, the heat generating system to control temperature, and the prototype structure. All parts and prototype kits are modeled by using
3D printing technology. The electrical circuit system developed and employed in the prototype kit could generate the heat to control the required
temperature between 36.5 °C and 37.5 °C, which is the optimum temperature for cull culture. Compared to the standard equipment, our portable
prototype kit yielded good diagnostic values (80% sensitivity, 100% specificity, and 90% accuracy), which are satisfactory and acceptable as a
practical tool to identify the culprit drugs.
© 2020 The Authors. Published by Elsevier Ltd.
This is an open access article under the CC BY-NC-ND license https://creativecommons.org/licenses/by-nc-nd/4.0/)
Peer-review under responsibility of the scientific committee of the FAIM 2020.
Keywords: allergy testing, drug allergy, ELISpot, 3D printing, portable.
1. Introduction
Drug allergies are types of adverse drug reactions due to the
unpredictable immunological responses to drugs [1]. Most of
drug allergic reactions have non-immediate onset and range
from mild to severe or even life threatening, depending on the
magnitude of cellular immune responses to the medications that
get into the body. White blood cells, particularly T-
lymphocytes, play a major role in the pathogenesis of drug-
induced non-immediate reactions by recognizing and reacting
to the administered drugs in susceptible patients [2]. Interferon
gamma (IFN-) is among the cytokines commonly secreted
from T-lymphocytes when drug allergic patients are exposed to
the allergic drugs. The measurement of drug-induced IFN-
releasing cells by using the Enzyme-Linked Immunosorbent
Spot technique (ELISpot) has been reported as a suitable tool
for drug allergy diagnosis [3].
IFN- ELISpot assay is sensitive to identify the culprit drugs
but required a special technique [3]. This technique involves 12
steps in conventional assay and requires specialized and
expensive equipment such as carbon dioxide (CO2) incubators,
CO2 tank and blood cell counting devices. There are only few
hospitals in Thailand with sufficient laboratory capacity to
perform ELISpot assay and most of the available equipment are
not movable. Therefore, it is currently not possible to perform
Available online at www.sciencedirect.com
ScienceDirect�
Procedia Manufacturing 00 (2021) 000–000
www.elsevier.com/locate/procedia
2351-9789 © 2020 The Authors. Published by Elsevier Ltd.
This is an open access article under the CC BY-NC-ND license https://creativecommons.org/licenses/by-nc-nd/4.0/)
Peer-review under responsibility of the scientific committee of the FAIM 2020.
30th International Conference on Flexible Automation and Intelligent Manufacturing (FAIM2021)
15-18 June 2021, Athens, Greece.
Development of Prototype Kit for Portable Drug Allergy Testing
Somkiat Tangjitsitcharoen1*, Jettanong Klaewsongkram2,
Angsumalin Senjuntichai1, Supranee Buranapraditkun2, Sutthinee Thongnop1
1Department of Industrial Engineering, Faculty of Engineering, Chulalongkorn University, Thailand
2Department of Medicine, Faculty of Medicine, Chulalongkorn University, Thailand
* Corresponding author. Tel.: +6-689-388-1688. E-mail address: somkiat.ta@eng.chula.ac.th
Abstract
The prototype kit for portable drug allergy testing is proposed and developed for easy-to-use in provincial hospitals where the standard equipment
for in vitro drug allergy diagnosis is still lacking. The Enzyme-Linked Immunosorbent Spot (ELISpot) technique is adopted for this purpose due
to its high sensitivity and specificity to detect cytokine-releasing cells, which is precise and suitable for non-instantaneously allergic tests. A
developed prototype for drug allergy testing has been considered in various aspects of the three main designs, comprising of the proposed testing
procedure kit, the heat generating system to control temperature, and the prototype structure. All parts and prototype kits are modeled by using
3D printing technology. The electrical circuit system developed and employed in the prototype kit could generate the heat to control the required
temperature between 36.5 °C and 37.5 °C, which is the optimum temperature for cull culture. Compared to the standard equipment, our portable
prototype kit yielded good diagnostic values (80% sensitivity, 100% specificity, and 90% accuracy), which are satisfactory and acceptable as a
practical tool to identify the culprit drugs.
© 2020 The Authors. Published by Elsevier Ltd.
This is an open access article under the CC BY-NC-ND license https://creativecommons.org/licenses/by-nc-nd/4.0/)
Peer-review under responsibility of the scientific committee of the FAIM 2020.
Keywords: allergy testing, drug allergy, ELISpot, 3D printing, portable.
1. Introduction
Drug allergies are types of adverse drug reactions due to the
unpredictable immunological responses to drugs [1]. Most of
drug allergic reactions have non-immediate onset and range
from mild to severe or even life threatening, depending on the
magnitude of cellular immune responses to the medications that
get into the body. White blood cells, particularly T-
lymphocytes, play a major role in the pathogenesis of drug-
induced non-immediate reactions by recognizing and reacting
to the administered drugs in susceptible patients [2]. Interferon
gamma (IFN-) is among the cytokines commonly secreted
from T-lymphocytes when drug allergic patients are exposed to
the allergic drugs. The measurement of drug-induced IFN-
releasing cells by using the Enzyme-Linked Immunosorbent
Spot technique (ELISpot) has been reported as a suitable tool
for drug allergy diagnosis [3].
IFN- ELISpot assay is sensitive to identify the culprit drugs
but required a special technique [3]. This technique involves 12
steps in conventional assay and requires specialized and
expensive equipment such as carbon dioxide (CO2) incubators,
CO2 tank and blood cell counting devices. There are only few
hospitals in Thailand with sufficient laboratory capacity to
perform ELISpot assay and most of the available equipment are
not movable. Therefore, it is currently not possible to perform
976 Somkiat Tangjitsitcharoen et al. / Procedia Manufacturing 51 (2020) 975–980
2 T. Somkiat et al. / Procedia Manufacturing 00 (2021) 000–000
drug allergy testing based on this technique in provincial
hospitals or any other remote areas.
3D printing technology is a technique for constructing
virtual reality structural parts in three dimensions. The
structural parts are designed through a computer program.
Therefore, 3D printing is able to form the structural parts of the
prototype kit with complexity according to the design. The
obtained prototype has the high strength as the desired design
[4-5].
Cell culture conditions inside the prototype kit require tight
temperature control between 36°C to 38°C to ensure the
optimum cell growth. In the prototype kit, there is an electric
circuit to maintain the internal environment to have the
temperature within the appropriate range. The circuit system
uses a small light bulb as a heat source with heat radiation and
electronic devices to control temperature inside the device
within the appropriate temperature range.
Hence, the purpose of this research is to develop a prototype
kit for portable drug allergy testing that is easy to use while still
maintaining a good quality compared to the standard method.
The prototype kit is supposed to be used in provincial hospitals
where the standard equipment for drug allergy testing are not
yet available.
2. Proposed testing procedures
The testing procedure of the drug allergy utilizing the
ELISpot technique in the prototype kit is proposed and
designed to reduce the testing process and shorten the testing
time but still be able to perform the test efficiently. Normally,
the cell culture process in the laboratory has used R10 medium
(Roswell Park Memorial Institute (RPMI1640) supplemented
with 100 U/ml of penicillin, 100 U/ml of streptomycin and 10%
heat-inactivated fetal bovine serum (FBS)) which must be
tested under 37°C and 5% CO2 condition. However, the tests in
the prototype kit will apply L15-10 medium (Leibovitz’s L-15
Medium (L15) supplemented with 100 U/ml of penicillin, 100
U/ml of streptomycin and 10% FBS) in cell culture to avoid the
use of carbon dioxide and control of temperature in the test [6].
In preparation process, the human IFN-γ antibody pre-coated
plate is employed to reduce testing time and errors that may be
caused during testing preparation. The signal amplification
process uses reagents mixed between second-antibody and
streptavidin-alkaline phosphatase (streptavidin-ALP). There is
non-chemical reaction to each other when mixing the two
solutions together. The procedures of drug allergy testing using
the ELISpot technique in the prototype kit can reduce the
testing process from 12 steps to 7 steps and reduce the testing
time approximately from 56 hours to 43 hours.
3. Temperature control
The design of electrical circuit system in the prototype kit is
taken into consideration for the optimum condition of cell
growth, which can generate the heat to control the suitable
temperatures from 36.5°C to 37.5°C that are the same as in the
laboratory. Circuit design uses the electricity or batteries as an
electrical power source which is converted into the prototype
kit as direct current (DC) of 5 volts. Hence, a 4.8 volt light bulb
is selected to generate the heat. The temperature control is set
if the temperature inside the kit is equal or lower than to 36.5°C,
the lamp will generate the heat in the kit until 37.5°C, and hence
the circuit will cut off the power to maintain the temperature
inside the kit, otherwise the cells will die.
The position of the lamp in the kit will affect the heat
distribution to the test wells. Hence, the heat distribution from
the lamp on the test well area is simulated and the heat radiation
is analyzed in the direction of top and right of the test well area
[7-8]. The simulation conditions are set as the ambient
temperature of 37°C, the emissivity is 0.8 and the free
convection coefficient in the air is 5 W/m2K by dividing the
mesh area at the highest resolution in order to study the heat
and temperature distribution characteristics at the test well and
determine the suitable direction to install the lamp in the
prototype kit [9].
The temperature control system is designed by using 3
incandescent light bulbs of 4.8 volts connected parallelly as
shown in Fig. 1.
Fig.1 Illustration of heat generation and temperature control system in a
prototype kit.
4. Design of prototype kit
In order to design the prototype kit structure and parts of the
prototype kit, it must correspond with the mechanism of drug
allergy testing using the ELISpot technique and design to
support the installation of electronic devices. The parts are
formed using 3D printing technology using photopolymer resin
materials.
5. Experimental setup and equipment
The sensor used in the prototype kit is infrared sensor that
can measure temperature and humidity in the air with the high
sensitivity and resolution of 0.1°C. The sensor used to measure
temperature in the laboratory is bimetallic type, which is stable,
and the resolution of temperature is 0.5°C. Therefore, the
calibration of the infrared sensor used in the prototype kit with
the temperature sensor used in the laboratory has been
compared and proved that the temperature around test well area
occurs within the range of 36.5°C to 37.5°C by referring to both
sensors.
In the prototype kit, the cell culture will be done by using
L15-10 medium and in the laboratory using R10 medium by
testing the cell culture in the positive wells and negative wells
using the same set of cells. After that, the spot number from the
T. Somkiat et al. / Procedia Manufacturing 00 (2021) 000–000 3
test is analyzed by statistical test using Wilcoxon Signed Rank
Test in GraphPad Prism 6.0 at the significance level of 0.05 in
order to consider the significant difference of the testing results
from the prototype kit and the laboratory in the positive wells
and negative wells.
The preliminary testing of the drug allergy prototype kit by
using the ELISpot technique is to assess the quality, the
reliability and the accuracy of the prototype kit. Ten
participants with drug allergy were tested in this study. Four
drugs which are anticonvulsant drugs, sulfa drugs, gouty
arthritis drugs and tuberculosis drugs are found common in
Thais drug allergy participants [10]. Two concentration of each
drug type were tested. Anticonvulsant drugs (phenytoin, at
concentrations of 20 µg/ml and 100 µg/ml), sulfa drugs
(bactrim, at concentrations of 40 µg/ml and 200 µg/ml), gout
drugs (oxypurinol, at concentrations of 10 µg/ml and 100
µg/ml) and tuberculosis drugs (IRZE, at concentrations of 10
µg/ml and 100 µg/ml) were added for ELISpot testing. This
study was approved by the Institutional Review Board of the
Faculty of Medicine, Chulalongkorn University (IRB No
724/62), and was conducted in accordance with the principles
of the Helsinki declaration ethical standards. All participants
provided written informed consent before the start of the study.
The procedure of drug allergy testing by using ELISpot
technique in the prototype kit are as follows:
Step 1: Add 100 µl of Peripheral Blood Mononuclear Cells
(PBMCs, 2.5x10
6
cells/ml in L15-10) into the drug-filled pre-
coated wells, negative well and positive wells. Incubate for 40
hours in a prototype with a temperature of 37°C. Do not move
the prototype in this time.
Step 2: Wash 6 times with PBS, 200 µl/well to remove the
cells.
Step 3: Add the mixed solution of second-antibody and
streptavidin-ALP, 100 µl/well and incubate for 2 hours at room
temperature
Step 4: Wash 6 times with PBS, 200 µl/well.
Step 5: Add substrate, 100 µl/well. The blue spots were
developed by leaving wells for 5-10 minutes at room
temperature.
Step 6: Wash 6 times with clean water and leave the wells
to dry.
Step 7: The spots were counted using automated ELISpot
reader (Immunospot analyzer, USA).
Normally, the cut-off in healthy control of phenytoin,
bactrim, oxypurinol and IRZE are 20 spots/million cells.
Results are expressed as the number of spots/million cells with
the drug, subtracted the spots from the cells incubated without
the drug. Consideration of the positive responses if the spot
numbers was greater than or equal to 20 spots/million cells.
6. Experimental results and discussions
To compare cell culture medium of R10 and L15-10, they
were tested with the same set of PBMCs for 5 experiments (Fig.
2 and Tables 1 and 2).
Experiment 1 Experiment 2 Experiment 3
R10 L15-10 R10 L15-10 R10 L15-10
Experiment 4 Experiment 5
R10 L15-10 R10 L15-10
Fig.2 Positive wells from testing cell culture mediums of R10 and L15-10.
Table 1 Number of spots in the positive wells from testing of cell culture
mediums of R10 and L15-10.
Experiment
Testing results in positive well
(number of spots/million cells)
R10 medium L15-10 medium
1 5052 5500
2 1660 1688
3 1644 1864
4 1596 1600
5 2492 2688
Median 1660 1864
From an analysis of the testing data in positive wells, the
R10 and L15-10 mediums were not different based on the
Wilcoxon Signed Rank Test (p-value > 0.05). It is indicated
that the prototype kit using L15-10 medium under the
temperature of 37°C was not different significantly from cell
culture tests using R10 medium at temperature control of 37°C
with 5% CO
2
as shown in Fig. 2 and Table 1. Hence, the drug
allergy testing using an ELISpot technique in the prototype kit
can be applied by using L15-10 medium.
Table 2 Number of spots in the negative wells from testing of cell culture
mediums of R10 and L15-10.
Experiment
Testing results in negative well
(number of spots/million cells)
R10 medium L15-10 medium
1 0 0
2 0 0
3 0 0
4 0 0
5 0 0
Median 0 0
In addition, we also found the data in negative wells were
not different between using of R10 and L15-10 mediums based
on the Wilcoxon Signed Rank Test (p-value > 0.05) in Table
2. This data suggested that the prototype kit can be applied by
using L15-10 medium under the temperature condition of 37°C
and clear background. It can be concluded that the number of
Somkiat Tangjitsitcharoen et al. / Procedia Manufacturing 51 (2020) 975–980 977
T. Somkiat et al. / Procedia Manufacturing 00 (2021) 000–000 3
test is analyzed by statistical test using Wilcoxon Signed Rank
Test in GraphPad Prism 6.0 at the significance level of 0.05 in
order to consider the significant difference of the testing results
from the prototype kit and the laboratory in the positive wells
and negative wells.
The preliminary testing of the drug allergy prototype kit by
using the ELISpot technique is to assess the quality, the
reliability and the accuracy of the prototype kit. Ten
participants with drug allergy were tested in this study. Four
drugs which are anticonvulsant drugs, sulfa drugs, gouty
arthritis drugs and tuberculosis drugs are found common in
Thais drug allergy participants [10]. Two concentration of each
drug type were tested. Anticonvulsant drugs (phenytoin, at
concentrations of 20 µg/ml and 100 µg/ml), sulfa drugs
(bactrim, at concentrations of 40 µg/ml and 200 µg/ml), gout
drugs (oxypurinol, at concentrations of 10 µg/ml and 100
µg/ml) and tuberculosis drugs (IRZE, at concentrations of 10
µg/ml and 100 µg/ml) were added for ELISpot testing. This
study was approved by the Institutional Review Board of the
Faculty of Medicine, Chulalongkorn University (IRB No
724/62), and was conducted in accordance with the principles
of the Helsinki declaration ethical standards. All participants
provided written informed consent before the start of the study.
The procedure of drug allergy testing by using ELISpot
technique in the prototype kit are as follows:
Step 1: Add 100 µl of Peripheral Blood Mononuclear Cells
(PBMCs, 2.5x10
6
cells/ml in L15-10) into the drug-filled pre-
coated wells, negative well and positive wells. Incubate for 40
hours in a prototype with a temperature of 37°C. Do not move
the prototype in this time.
Step 2: Wash 6 times with PBS, 200 µl/well to remove the
cells.
Step 3: Add the mixed solution of second-antibody and
streptavidin-ALP, 100 µl/well and incubate for 2 hours at room
temperature
Step 4: Wash 6 times with PBS, 200 µl/well.
Step 5: Add substrate, 100 µl/well. The blue spots were
developed by leaving wells for 5-10 minutes at room
temperature.
Step 6: Wash 6 times with clean water and leave the wells
to dry.
Step 7: The spots were counted using automated ELISpot
reader (Immunospot analyzer, USA).
Normally, the cut-off in healthy control of phenytoin,
bactrim, oxypurinol and IRZE are 20 spots/million cells.
Results are expressed as the number of spots/million cells with
the drug, subtracted the spots from the cells incubated without
the drug. Consideration of the positive responses if the spot
numbers was greater than or equal to 20 spots/million cells.
6. Experimental results and discussions
To compare cell culture medium of R10 and L15-10, they
were tested with the same set of PBMCs for 5 experiments (Fig.
2 and Tables 1 and 2).
Experiment 1 Experiment 2 Experiment 3
R10 L15-10 R10 L15-10 R10 L15-10
Experiment 4 Experiment 5
R10 L15-10 R10 L15-10
Fig.2 Positive wells from testing cell culture mediums of R10 and L15-10.
Table 1 Number of spots in the positive wells from testing of cell culture
mediums of R10 and L15-10.
Experiment
Testing results in positive well
(number of spots/million cells)
R10 medium L15-10 medium
1 5052 5500
2 1660 1688
3 1644 1864
4 1596 1600
5 2492 2688
Median 1660 1864
From an analysis of the testing data in positive wells, the
R10 and L15-10 mediums were not different based on the
Wilcoxon Signed Rank Test (p-value > 0.05). It is indicated
that the prototype kit using L15-10 medium under the
temperature of 37°C was not different significantly from cell
culture tests using R10 medium at temperature control of 37°C
with 5% CO
2
as shown in Fig. 2 and Table 1. Hence, the drug
allergy testing using an ELISpot technique in the prototype kit
can be applied by using L15-10 medium.
Table 2 Number of spots in the negative wells from testing of cell culture
mediums of R10 and L15-10.
Experiment
Testing results in negative well
(number of spots/million cells)
R10 medium L15-10 medium
1 0 0
2 0 0
3 0 0
4 0 0
5 0 0
Median 0 0
In addition, we also found the data in negative wells were
not different between using of R10 and L15-10 mediums based
on the Wilcoxon Signed Rank Test (p-value > 0.05) in Table
2. This data suggested that the prototype kit can be applied by
using L15-10 medium under the temperature condition of 37°C
and clear background. It can be concluded that the number of
978 Somkiat Tangjitsitcharoen et al. / Procedia Manufacturing 51 (2020) 975–980
4 T. Somkiat et al. / Procedia Manufacturing 00 (2021) 000–000
spots in the positive wells are actually from IFN- secretion of
stimulated cells. The prototype kit is developed in sterilizing
condition because no found contamination during testing.
Figure 3 shows that the proposed circuit can generate the
heat in the prototype kit at the desired temperature of 36.5°C to
37.5°C and the temperature control system can maintain the
temperature inside the prototype kit throughout 48 hours. The
experimental results can confirm that it is possible to control
and maintain the desired temperature under a specified range in
the prototype kit completely.
However, using a power bank as a power supply for the
prototype kit, it was found that the prototype kit could be
operated by using two power banks for the entire 48-hour
testing period. The first power bank will run out at 25 hours of
operation and the second one will replace until the end of
testing period.
Fig.3 Illustration of relation between temperature and time in the prototype kit.
Comparing the testing operation of the prototype kit
between using power bank as a power source and using the
house electricity as a power source, it is found that the power
bank will take time (~ 1 hour) to increase the initial temperature
to 36.5°C, which is longer than using the house electricity (~
2.5 hours). Because the power bank is the direct current
electricity that has a voltage of 5 to 12 volts, while the home
electricity is alternating current with a voltage of 220 volts.
When using the home electricity as a power source, it can
supply electricity to light bulbs with more voltages than the
power bank. Light bulbs that use the house electricity as a
power source will be brighter and can distribute more heat to
the surrounding area. The brighter lamp will be able to radiate
more heat, it takes less time from the initial temperature to
36.5°C than using a power bank as a power source. Therefore,
the prototype kit can even use power bank as a power source,
but it may take more time to the desired temperature.
The heat distribution from the lamp radiation using a
computer simulation illustrates that the heat dissipation occurs
at the test wells from the top direction with the different surface
colors corresponding to the different temperatures as shown in
Fig.4.
It was found that the test wells area shows the red color in
the middle and slightly become to the green color in the edge.
The green color represents the temperature of 37.34°C while
the red color is 37.77°C. The simulation shows a good and
consistent heat distribution in the test well strip. It is expected
that the cell culture in each well can grow well in every well
and the number of spots will be correct and consistent.
Fig.4 Illustration of heat distribution and radiation of lamp from top direction
of test wells.
The structural design and dimension of prototype kit is
modeled and developed using computer programs. The
prototype kit in isometric view is 96 mm x 151 mm x 89 mm
(width x length x height) as shown in Fig.5. The parts of
prototype kit are formed utilizing the resin material employing
by the 3D printing technology as shown in Fig.6.
Fig.5 Dimension of a prototype kit in isometric view.
Fig.6 Illustration of prototype kit formed by 3D printing technology.
89 mm
T. Somkiat et al. / Procedia Manufacturing 00 (2021) 000–000 5
The prototype kit for drug allergy testing is proved with an
ELISpot technique, which is conducted parallelly with the
laboratory tests by using the same cell culture in positive and
negative wells.
It is clear that the prototype kit can test the drug allergy by
using ELISpot technique. The result in the negative and
positive wells are not different between the prototype kit and
the conventional method in the laboratory (p > 0.05). It is
confirmed that the procedures and conditions for drug allergy
testing of the prototype kit are consistent and equivalent with
laboratory tests referring to the testing results.
To evaluate the prototype kit, 10 participants with drug
allergy were tested in this study by using ELISpot assay with 4
drugs. The experimentally obtained results from the prototype
kit are illustrated in Table 3.
Table 3 Number of spots obtained from prototype kit and laboratory tests.
Experim
ent
Drug
type
Test
by
Number of spots/million cells
Positi
ve
Negati
ve
Low
concentra
tion drug
High
concentra
tion drug
1 Phenyto
in
Lab 484 0 0 0
Kit 484 0 0 16
2 Bactrim Lab 1716 0 216 356
Kit 1696 0 216 352
3 Bactrim Lab 1708 8 0 8
Kit 1716 8 0 0
4 Bactrim Lab 1568 0 372 612
Kit 1572 0 372 616
5 Oxypuri
nol
Lab 1596 0 0 0
Kit 1600 0 0 0
6 Oxypuri
nol
Lab 412 0 44 68
Kit 408 0 12 16
7 Oxypuri
nol
Lab 808 0 0 72
Kit 816 0 60 48
8 IRZE Lab 252 0 0 0
Kit 268 0 0 0
9 IRZE Lab 484 0 48 52
Kit 496 4 28 20
10 IRZE Lab 1656 0 0 0
Kit 1652 0 0 0
Referring to statistical tests at significance level of 0.05, the
P-value of Wilcoxon Signed Rank Test is 0.3438, which
exceeds 0.05. It means that the prototype kit can be used to test
the drug allergy. The spots in not only positive wells but also
negative wells are not significantly different from laboratory
tests. It is indicated that the drug allergy test in the prototype
kit is consistent with the laboratory tests which are equivalent.
The calculation of sensitivity, specificity, accuracy and
predictive values of prototype kit is shown in Table 4.
Table 4 Results of drug allergy testing using prototype kit with ELISpot
technique compared to laboratory tests.
Drug allergy testing results of
laboratory
Positive result
(+)
Negative result
) - (
Drug allergy
testing results of
prototype kit
Positive result
(+) 4 (a) 0 (b)
Negative
result )- ( 1 (c) 5 (d)
sensitivity is a/a+c = 80%
specificity is d/b+d = 100%
accuracy is [a+d]/[a+b+c+d] = 90%
positive predictive value is a/a+b = 100%
negative predictive value is d/c+d = 83%
Evaluation of drug allergy testing using the prototype kit
comparing to the laboratory tests, it is found that the prototype
kit has sensitivity of 80%, specificity of 100%, accuracy of
90%, and positive predictive value of 100% and negative
predictive value of 83%. It can be seen that the experimental
results have the high reliable level, which imply that the
portable drug allergy prototype kit is promising and able to be
tested for drug allergy using the ELISpot technique which is
equivalent to the laboratory tests.
7. Conclusions
Prototype kit has been developed for drug allergy testing
using the ELISpot technique. The drug allergy testing process
has been reduced within the shortage time. The L15-10 cell
culture mediums in the prototype kit is proposed and tested
under a controlled condition of 37°C. The prototype kit can use
both electricity and power bank as power sources. The resin
material with 3D printing technology is utilized to form the
structures and parts of the prototype kit. The drug allergy
testing has been applied to prove the prototype kit with 4 drugs,
which are anticonvulsant drugs, sulfa drugs, gouty arthritis
drugs and tuberculosis drugs. It was found that drug allergy
testing results in positive and negative wells have P-value
greater than 0.05 in every test. It is indicated that the drug
allergy testing results using the ELISpot technique in the
prototype kit can provide non-significantly different results
from the laboratory tests.
Evaluation of drug allergy testing using the prototype kit has
been compared with the laboratory test. It is proved that the
prototype kit has sensitivity of 80%, specificity of 100%,
accuracy of 90%, and positive predictive value of 100% and
negative predictive value of 83%. It could be concluded that
the drug allergy testing using the ELISpot technique with the
prototype kit is reliable and equivalent to the laboratory tests.
Acknowledgements
This work was performed by the funding of
Ratchadaphiseksomphot Endowment Fund, Chulalongkorn
University, 2019 and Chulalongkorn Medical Research Center
(Chula MRC), Department of Medicine, Faculty of Medicine,
Somkiat Tangjitsitcharoen et al. / Procedia Manufacturing 51 (2020) 975–980 979
T. Somkiat et al. / Procedia Manufacturing 00 (2021) 000–000 5
The prototype kit for drug allergy testing is proved with an
ELISpot technique, which is conducted parallelly with the
laboratory tests by using the same cell culture in positive and
negative wells.
It is clear that the prototype kit can test the drug allergy by
using ELISpot technique. The result in the negative and
positive wells are not different between the prototype kit and
the conventional method in the laboratory (p > 0.05). It is
confirmed that the procedures and conditions for drug allergy
testing of the prototype kit are consistent and equivalent with
laboratory tests referring to the testing results.
To evaluate the prototype kit, 10 participants with drug
allergy were tested in this study by using ELISpot assay with 4
drugs. The experimentally obtained results from the prototype
kit are illustrated in Table 3.
Table 3 Number of spots obtained from prototype kit and laboratory tests.
Experim
ent
Drug
type
Test
by
Number of spots/million cells
Positi
ve
Negati
ve
Low
concentra
tion drug
High
concentra
tion drug
1 Phenyto
in
Lab 484 0 0 0
Kit 484 0 0 16
2 Bactrim Lab 1716 0 216 356
Kit 1696 0 216 352
3 Bactrim Lab 1708 8 0 8
Kit 1716 8 0 0
4 Bactrim Lab 1568 0 372 612
Kit 1572 0 372 616
5 Oxypuri
nol
Lab 1596 0 0 0
Kit 1600 0 0 0
6 Oxypuri
nol
Lab 412 0 44 68
Kit 408 0 12 16
7 Oxypuri
nol
Lab 808 0 0 72
Kit 816 0 60 48
8 IRZE Lab 252 0 0 0
Kit 268 0 0 0
9 IRZE Lab 484 0 48 52
Kit 496 4 28 20
10 IRZE Lab 1656 0 0 0
Kit 1652 0 0 0
Referring to statistical tests at significance level of 0.05, the
P-value of Wilcoxon Signed Rank Test is 0.3438, which
exceeds 0.05. It means that the prototype kit can be used to test
the drug allergy. The spots in not only positive wells but also
negative wells are not significantly different from laboratory
tests. It is indicated that the drug allergy test in the prototype
kit is consistent with the laboratory tests which are equivalent.
The calculation of sensitivity, specificity, accuracy and
predictive values of prototype kit is shown in Table 4.
Table 4 Results of drug allergy testing using prototype kit with ELISpot
technique compared to laboratory tests.
Drug allergy testing results of
laboratory
Positive result
(+)
Negative result
) - (
Drug allergy
testing results of
prototype kit
Positive result
(+) 4 (a) 0 (b)
Negative
result )- ( 1 (c) 5 (d)
sensitivity is a/a+c = 80%
specificity is d/b+d = 100%
accuracy is [a+d]/[a+b+c+d] = 90%
positive predictive value is a/a+b = 100%
negative predictive value is d/c+d = 83%
Evaluation of drug allergy testing using the prototype kit
comparing to the laboratory tests, it is found that the prototype
kit has sensitivity of 80%, specificity of 100%, accuracy of
90%, and positive predictive value of 100% and negative
predictive value of 83%. It can be seen that the experimental
results have the high reliable level, which imply that the
portable drug allergy prototype kit is promising and able to be
tested for drug allergy using the ELISpot technique which is
equivalent to the laboratory tests.
7. Conclusions
Prototype kit has been developed for drug allergy testing
using the ELISpot technique. The drug allergy testing process
has been reduced within the shortage time. The L15-10 cell
culture mediums in the prototype kit is proposed and tested
under a controlled condition of 37°C. The prototype kit can use
both electricity and power bank as power sources. The resin
material with 3D printing technology is utilized to form the
structures and parts of the prototype kit. The drug allergy
testing has been applied to prove the prototype kit with 4 drugs,
which are anticonvulsant drugs, sulfa drugs, gouty arthritis
drugs and tuberculosis drugs. It was found that drug allergy
testing results in positive and negative wells have P-value
greater than 0.05 in every test. It is indicated that the drug
allergy testing results using the ELISpot technique in the
prototype kit can provide non-significantly different results
from the laboratory tests.
Evaluation of drug allergy testing using the prototype kit has
been compared with the laboratory test. It is proved that the
prototype kit has sensitivity of 80%, specificity of 100%,
accuracy of 90%, and positive predictive value of 100% and
negative predictive value of 83%. It could be concluded that
the drug allergy testing using the ELISpot technique with the
prototype kit is reliable and equivalent to the laboratory tests.
Acknowledgements
This work was performed by the funding of
Ratchadaphiseksomphot Endowment Fund, Chulalongkorn
University, 2019 and Chulalongkorn Medical Research Center
(Chula MRC), Department of Medicine, Faculty of Medicine,
980 Somkiat Tangjitsitcharoen et al. / Procedia Manufacturing 51 (2020) 975–980
6 T. Somkiat et al. / Procedia Manufacturing 00 (2021) 000–000
Chulalongkorn University, Bangkok, Thailand. This work was
also supported by Division of Allergy and Clinical
Immunology, Department of Medicine, Faculty of Medicine,
Chulalongkorn University, Bangkok, Thailand and Center of
Excellence in Vaccine Research and Development (Chula
Vaccine Research Center-Chula VRC), Faculty of Medicine,
Chulalongkorn University, Bangkok 10330, Thailand.
References
[1] Suwankesawong W, Sriphiromya P, Tragulpiankit P, Phetcharat C,
Sornsrivichai V. Evaluation of Thai Algorithm Usage for Adverse Drug
Reaction Monitoring. Journal of Health Science 2016;25:4.
[2] Malcolm R. A., Richard P, Stuart F, Nicholas A. W. An introduction to
stem cells. Journal of Pathology 2002;197:419-423.
[3] Tanvarasethee B. Skin testing and cytokine secretion from drug specific
peripheral blood mononuclear cell using elispot assay in the diagnosis of
nonimmediate reaction to cephalosporin. Dissertation, Chulalongkorn
University; 2010.
[4] Tangjitsitcharoen S, Khomvilai S, Rattanatraiphob M et al. Development
of prototype of automated hemorrhoid ligator. Journal of Science and
Technology, Srinakharinwirot University 2017;9:179-187.
[5] Ren X, Shao H, Lin T et al. 3D gel-printing—An additive manufacturing
method for producing complex shape parts. Journal of Materials and
Design 2016;101:80-87.
[6] Thermo Fisher (2019) Leibovitz's L-15 Medium.
https://www.thermofisher.com/order/catalog/product/ Leibovitz's L-15
Medium.
Accessed 26 November 2019
[7] Padungyat T. Turbulent convection in triangular duct with constant heat
flux. Dissertation, Chulalongkorn University; 1997.
[8] Howell JR, Menguc MP, Siegel R (2015) Thermal radiation heat transfer,
6th edn. New York.
[9] Nilpueng K. Heat Transfer Coefficient, Pressure Drop and Flow Patterns
of Water-Air Two-Phase Flow Through a Plate Heat Exchanger.
Dissertation, King Mongkut's University of Technology Thonburi; 2005.
[10] Klaewsongkram J, Sukasem C, Thantiworasit P et al. Analysis of HLA-B
Allelic Variation and IFN-γ ELISpot Responses in Patients with Severe
Cutaneous Adverse Reactions Associated with Drugs. J Allergy Clin
Immunol Pract 2019;7(1): 219-227.
