https://orcid.org/0009-0008-7983-4185
Abstract
For individuals with infectious diseases, early and accurate diagnosis is critical. A rapid diagnosis allows for prompt and effective treatment and increases the chance of a full recovery without complications. Additionally, when containing a wide-scale infectious disease outbreak, circumstances are significantly improved by the ability to test the populace frequently, swiftly, and affordably. Regarding specificity and sensitivity, nucleic acid amplification tests (NAAT) are one of the best options for diagnosing infectious diseases. Historically, polymerase chain reaction (PCR) has been used, but complex thermocycling and complicated PCR protocols have often limited PCR to clinical settings. Due to increased simplicity, the isothermal NAAT recombinase polymerase amplification (RPA) has the potential to deliver reliable Point-of-Care (POC) diagnostics in low-resource settings. When designing POC devices for isothermal NAATs, creating isothermal temperature conditions is perhaps the most significant challenge. This work presents a flexible and robust device capable of incubating 3 RPA reactions for simultaneous amplification in conditions conducive to POC testing. The device costs ∼$60 USD to construct and is easy to assemble. A battery-powered polyimide thin-film resistive heater provides energy, and the device only requires power for a fraction of the total incubation time. The device uses a phase change material (PCM) to regulate temperature to avoid the complexity of a microcontroller. RPA reactions were successfully incubated for 30 min using the device.
References
1.
Hwang
,
H.
,
Hwang
,
B.-Y.
, and
Bueno
,
J.
, 2018
, “
Biomarkers in Infectious Diseases
,” Dis. Markers
,
2018
, p. 8509127
.10.1155/2018/85091272.
Ritchie
,
H.
, and
Roser
,
M.
, 2018
, Causes of Death
,
Our World Data
, Oxford, UK.3.
Chen
,
H.
,
Liu
,
K.
,
Li
,
Z.
, and
Wang
,
P.
, 2019
, “
Point of Care Testing for Infectious Diseases
,” Clin. Chim. Acta Int. J. Clin. Chem.
,
493
, pp. 138
–147
.10.1016/j.cca.2019.03.0084.
World Health Organization
, 2020
, “
The Top 10 Causes of Death
,” World Health Organization, Geneva, Switzerland, accessed May 10, 2021, https://www.who.int/news-room/fact-sheets/detail/the-top-10-causes-of-death5.
Suea-Ngam
,
A.
,
Bezinge
,
L.
,
Mateescu
,
B.
,
Howes
,
P. D.
,
deMello
,
A. J.
, and
Richards
,
D. A.
, 2020
, “
Enzyme-Assisted Nucleic Acid Detection for Infectious Disease Diagnostics: Moving Toward the Point-of-Care
,” ACS Sens.
,
5
(9
), pp. 2701
–2723
.10.1021/acssensors.0c014886.
World Health Organization, 2022, “
WHO Coronavirus (COVID-19) Dashboard
,” World Health Organization, Geneva, Switzerland, accessed July 20, 2022, https://covid19.who.int7.
Yager
,
P.
,
Domingo
,
G. J.
, and
Gerdes
,
J.
, 2008
, “
Point-of-Care Diagnostics for Global Health
,” Annu. Rev. Biomed. Eng.
,
10
(1
), pp. 107
–144
.10.1146/annurev.bioeng.10.061807.1605248.
Samuel
,
L.
, 2020
, “
Point-of-Care Testing in Microbiology
,” Clin. Lab. Med.
,
40
(4
), pp. 483
–494
.10.1016/j.cll.2020.08.0069.
Karaivanov
,
A.
,
Lu
,
S. E.
,
Shigeoka
,
H.
,
Chen
,
C.
, and
Pamplona
,
S.
, 2021
, “
Face Masks, Public Policies and Slowing the Spread of COVID-19: Evidence From Canada
,” J. Health Econ.
,
78
, p. 102475
.10.1016/j.jhealeco.2021.10247510.
Rutayisire
,
E.
,
Nkundimana
,
G.
,
Mitonga
,
H. K.
,
Boye
,
A.
, and
Nikwigize
,
S.
, 2020
, “
What Works and What Does Not Work in Response to COVID-19 Prevention and Control in Africa
,” Int. J. Infect. Dis.
,
97
, pp. 267
–269
.10.1016/j.ijid.2020.06.02411.
Hansen
,
G. T.
, 2020
, “
Point-of-Care Testing in Microbiology: A Mechanism for Improving Patient Outcomes
,” Clin. Chem.
,
66
(1
), pp. 124
–137
.10.1373/clinchem.2019.30478212.
Niezen
,
G.
,
Eslambolchilar
,
P.
, and
Thimbleby
,
H.
, 2016
, “
Open-Source Hardware for Medical Devices
,” BMJ Innov.
,
2
(2
), pp. 78
–83
.10.1136/bmjinnov-2015-00008013.
Wu
,
G.
, and
Zaman
,
M. H.
, 2012
, “
Low-Cost Tools for Diagnosing and Monitoring HIV Infection in Low-Resource Settings
,” Bull. World Health Organ.
,
90
(12
), pp. 914
–920
.10.2471/BLT.12.10278014.
Drain
,
P. K.
,
Hyle
,
E. P.
,
Noubary
,
F.
,
Freedberg
,
K. A.
,
Wilson
,
D.
,
Bishai
,
W.
,
Rodriguez
,
W.
, and
Bassett
,
I. V.
, 2014
, “
Evaluating Diagnostic Point-of-Care Tests in Resource-Limited Settings
,” Lancet Infect. Dis.
,
14
(3
), pp. 239
–249
.10.1016/S1473-3099(13)70250-015.
Mauk
,
M. G.
,
Song
,
J.
,
Liu
,
C.
, and
Bau
,
H. H.
, 2018
, “
Simple Approaches to Minimally-Instrumented, Microfluidic-Based Point-of-Care Nucleic Acid Amplification Tests
,” Biosensor
,
8
(1
), p. 17
.10.3390/bios801001716.
Obande
,
G. A.
, and
Banga Singh
,
K. K.
, 2020
, “
Current and Future Perspectives on Isothermal Nucleic Acid Amplification Technologies for Diagnosing Infections
,” Infect. Drug Resist.
,
13
, pp. 455
–483
.10.2147/IDR.S21757117.
Maffert
,
P.
,
Reverchon
,
S.
,
Nasser
,
W.
,
Rozand
,
C.
, and
Abaibou
,
H.
, 2017
, “
New Nucleic Acid Testing Devices to Diagnose Infectious Diseases in Resource-Limited Settings
,” Eur. J. Clin. Microbiol. Infect. Dis. Off. Publ. Eur. Soc. Clin. Microbiol.
,
36
(10
), pp. 1717
–1731
.10.1007/s10096-017-3013-918.
Craw
,
P.
, and
Balachandran
,
W.
, 2012
, “
Isothermal Nucleic Acid Amplification Technologies for Point-of-Care Diagnostics: A Critical Review
,” Lab. Chip
,
12
(14
), pp. 2469
–2486
.10.1039/c2lc40100b19.
Miralles
,
V.
,
Huerre
,
A.
,
Malloggi
,
F.
, and
Jullien
,
M.-C.
, 2013
, “
A Review of Heating and Temperature Control in Microfluidic Systems: Techniques and Applications
,” Diagnostics
,
3
(1
), pp. 33
–67
.10.3390/diagnostics301003320.
Li
,
J.
,
Macdonald
,
J.
, and
Stetten
,
F. V.
, 2019
, “
Review: A Comprehensive Summary of a Decade Development of the Recombinase Polymerase Amplification
,” Analyst
,
144
(1
), pp. 31
–67
.10.1039/C8AN01621F21.
Lobato
,
I. M.
, and
O'Sullivan
,
C. K.
, 2018
, “
Recombinase Polymerase Amplification: Basics, Applications and Recent Advances
,” TrAC Trends Anal. Chem.
,
98
, pp. 19
–35
.10.1016/j.trac.2017.10.01522.
Daher
,
R. K.
,
Stewart
,
G.
,
Boissinot
,
M.
, and
Bergeron
,
M. G.
, 2016
, “
Recombinase Polymerase Amplification for Diagnostic Applications
,” Clin. Chem.
,
62
(7
), pp. 947
–958
.10.1373/clinchem.2015.24582923.
Qiu
,
X.
,
Zhang
,
S.
,
Xiang
,
F.
,
Wu
,
D.
,
Guo
,
M.
,
Ge
,
S.
,
Li
,
K.
,
Ye
,
X.
,
Xia
,
N.
, and
Qian
,
S.
, 2017
, “
Instrument-Free Point-of-Care Molecular Diagnosis of H1N1 Based on Microfluidic Convective PCR
,” Sens. Actuators B Chem.
,
243
, pp. 738
–744
.10.1016/j.snb.2016.12.05824.
Abdullahi
,
U. F.
,
Naim
,
R.
,
Taib
,
W. R. W.
,
Saleh
,
A.
,
Muazu
,
A.
,
Baig
,
S. A.
, and
Baig
,
A. A.
, 2015
, “
Loop-Mediated Isothermal Amplification (LAMP), An Innovation in Gene Amplification: Bridging the Gap in Molecular Diagnostics; A Review
,” Indian J. Sci. Technol.
,
8
(17
), pp. 1
–12
.10.17485/ijst/2015/v8i17/5576725.
Piepenburg
,
O.
,
Williams
,
C. H.
,
Stemple
,
D. L.
, and
Armes
,
N. A.
, 2006
, “
DNA Detection Using Recombination Proteins
,” PLoS Biol.
,
4
(7
), p. e204
.10.1371/journal.pbio.004020426.
Buser
,
J. R.
,
Diesburg
,
S.
,
Singleton
,
J.
,
Guelig
,
D.
,
Bishop
,
J. D.
,
Zentner
,
C.
,
Burton
,
R.
,
LaBarre
,
P.
,
Yager
,
P.
, and
Weigl
,
B. H.
, 2015
, “
Precision Chemical Heating for Diagnostic Devices
,” Lab. Chip
,
15
(23
), pp. 4423
–4432
.10.1039/C5LC01053E27.
Huang
,
S.
,
Do
,
J.
,
Mahalanabis
,
M.
,
Fan
,
A.
,
Zhao
,
L.
,
Jepeal
,
L.
,
Singh
,
S. K.
, and
Klapperich
,
C. M.
, 2013
, “
Low Cost Extraction and Isothermal Amplification of DNA for Infectious Diarrhea Diagnosis
,” PLoS One
,
8
(3
), p. e60059
.10.1371/journal.pone.006005928.
Goertz
,
J. P.
,
Colvin
,
K. M.
,
Lippe
,
A. B.
,
Daristotle
,
J. L.
,
Kofinas
,
P.
, and
White
,
I. M.
, 2018
, “
Multistage Chemical Heating for Instrument-Free Biosensing
,” ACS Appl. Mater. Interfaces
,
10
(39
), pp. 33043
–33048
.10.1021/acsami.8b1161129.
Liao
,
S.-C.
,
Peng
,
J.
,
Mauk
,
M. G.
,
Awasthi
,
S.
,
Song
,
J.
,
Friedman
,
H.
,
Bau
,
H. H.
, and
Liu
,
C.
, 2016
, “
Smart Cup: A Minimally-Instrumented, Smartphone-Based Point-of-Care Molecular Diagnostic Device
,” Sens. Actuators B Chem.
,
229
, pp. 232
–238
.10.1016/j.snb.2016.01.07330.
Linnes
,
J. C.
,
Fan
,
A.
,
Rodriguez
,
N. M.
,
Lemieux
,
B.
,
Kong
,
H.
, and
Klapperich
,
C. M.
, 2014
, “
Paper-Based Molecular Diagnostic for Chlamydia Trachomatis
,” RSC Adv.
,
4
(80
), pp. 42245
–42251
.10.1039/C4RA07911F31.
Zhang
,
Y.
,
Zhang
,
L.
,
Sun
,
J.
,
Liu
,
Y.
,
Ma
,
X.
,
Cui
,
S.
,
Ma
,
L.
,
Xi
,
J. J.
, and
Jiang
,
X.
, 2014
, “
Point-of-Care Multiplexed Assays of Nucleic Acids Using Microcapillary-Based Loop-Mediated Isothermal Amplification
,” Anal. Chem.
,
86
(14
), pp. 7057
–7062
.10.1021/ac501433232.
Kenisarin
,
M. M.
, 2014
, “
Thermophysical Properties of Some Organic Phase Change Materials for Latent Heat Storage. A Review
,” Sol. Energy
,
107
, pp. 553
–575
.10.1016/j.solener.2014.05.00133.
Singleton
,
J.
,
Zentner
,
C.
,
Buser
,
J.
,
Yager
,
P.
,
LaBarre
,
P.
, and
Weigl
,
B. H.
, 2013
, “
Instrument-Free Exothermic Heating With Phase Change Temperature Control for Paper Microfluidic Devices
,” Proc. SPIE
,
8615
, p. 86150R
.10.1117/12.200592834.
Curtis
,
K. A.
,
Rudolph
,
D. L.
,
Morrison
,
D.
,
Guelig
,
D.
,
Diesburg
,
S.
,
McAdams
,
D.
,
Burton
,
R. A.
,
LaBarre
,
P.
, and
Owen
,
M.
, 2016
, “
Single-Use, Electricity-Free Amplification Device for Detection of HIV-1
,” J. Virol. Methods
,
237
, pp. 132
–137
.10.1016/j.jviromet.2016.09.00735.
Sema
,
M.
,
Alemu
,
A.
,
Bayih
,
A. G.
,
Getie
,
S.
,
Getnet
,
G.
,
Guelig
,
D.
,
Burton
,
R.
,
LaBarre
,
P.
, and
Pillai
,
D. R.
, 2015
, “
Evaluation of Non-Instrumented Nucleic Acid Amplification by Loop-Mediated Isothermal Amplification (NINA-LAMP) for the Diagnosis of Malaria in Northwest Ethiopia
,” Malar. J.
,
14
(1
), p. 44
.10.1186/s12936-015-0559-936.
Singleton
,
J.
,
Osborn
,
J. L.
,
Lillis
,
L.
,
Hawkins
,
K.
,
Guelig
,
D.
,
Price
,
W.
,
Johns
,
R.
,
Ebels
,
K.
,
Boyle
,
D.
,
Weigl
,
B.
, and
LaBarre
,
P.
, 2014
, “
Electricity-Free Amplification and Detection for Molecular Point-of-Care Diagnosis of HIV-1
,” PLoS One
,
9
(11
), p. e113693
.10.1371/journal.pone.011369337.
Song
,
J.
,
Mauk
,
M. G.
,
Hackett
,
B. A.
,
Cherry
,
S.
,
Bau
,
H. H.
, and
Liu
,
C.
, 2016
, “
Instrument-Free Point-of-Care Molecular Detection of Zika Virus
,” Anal. Chem.
,
88
(14
), pp. 7289
–7294
.10.1021/acs.analchem.6b0163238.
Vloemans
,
D.
,
Dal Dosso
,
F.
,
Lopez
,
C.
,
Macdonald
,
J.
, and
Lammertyn
,
J.
, 2019
, “
Phase Change Materials as an Enabler for Malaria Detection in Lowresource Settings
,” 23rd International Conference on Miniaturized Systems for Chemistry and Life Sciences
, Basel, Switzerland, Oct. 27
–31
.https://research.usc.edu.au/esploro/outputs/9948900760262139.
Vloemans
,
D.
,
Dal Dosso
,
F.
,
Verboven
,
P.
,
Nicolai
,
B.
, and
Lammertyn
,
J.
, 2020
, “
Exploiting Phase Change Materials in Tunable Passive Heating System for Low-Resource Point-of-Care Diagnostics
,” Appl. Therm. Eng.
,
173
, p. 115269
.10.1016/j.applthermaleng.2020.11526940.
Pardy
,
T.
,
Tulp
,
I.
,
Kremer
,
C.
,
Rang
,
T.
, and
Stewart
,
R.
, 2017
, “
Integrated Self-Regulating Resistive Heating for Isothermal Nucleic Acid Amplification Tests (NAAT) in Lab-on-a-Chip (LoC) Devices
,” PLoS One
,
12
(12
), p. e0189968
.10.1371/journal.pone.018996841.
Liu
,
C.
,
Mauk
,
M. G.
,
Hart
,
R.
,
Bonizzoni
,
M.
,
Yan
,
G.
, and
Bau
,
H. H.
, 2012
, “
A Low-Cost Microfluidic Chip for Rapid Genotyping of Malaria-Transmitting Mosquitoes
,” Plos One
,
7
(8
), p. e42222
.10.1371/journal.pone.004222242.
Pardy
,
T.
,
Rang
,
T.
, and
Tulp
,
I.
, 2017
, “
Development of Temperature Control Solutions for Non-Instrumented Nucleic Acid Amplification Tests (NINAAT)
,” Micromachines
,
8
(6
), p. 180
.10.3390/mi806018043.
Veltkamp
,
H.-W.
,
Akegawa Monteiro
,
F.
,
Sanders
,
R.
,
Wiegerink
,
R.
, and
Lötters
,
J.
, 2020
, “
Disposable DNA Amplification Chips With Integrated Low-Cost Heaters
,” Micromachines
,
11
(3
), p. 238
.10.3390/mi1103023844.
Pardy
,
T.
,
Sink
,
H.
,
Koel
,
A.
, and
Rang
,
T.
, 2019
, “
Development of a Low-Cost, Wireless Smart Thermostat for Isothermal DNA Amplification in Lab-On-A-Chip Devices
,” Micromachines
,
10
(7
), p. 437
.10.3390/mi1007043745.
Myers
,
F. B.
,
Henrikson
,
R. H.
,
Bone
,
J.
, and
Lee
,
L. P.
, 2013
, “
A Handheld Point-of-Care Genomic Diagnostic System
,” PLoS One
,
8
(8
), p. e70266
.10.1371/journal.pone.007026646.
Streit
,
P.
,
Nestler
,
J.
,
Shaporin
,
A.
,
Graunitz
,
J.
, and
Otto
,
T.
, 2018
, “
Design Methodology and Results Evaluation of a Heating Functionality in Modular Lab-on-Chip Systems
,” J. Micromech. Microeng.
,
28
(6
), p. 064001
.10.1088/1361-6439/aab0ca47.
Stedtfeld
,
R. D.
,
Tourlousse
,
D. M.
,
Seyrig
,
G.
,
Stedtfeld
,
T. M.
,
Kronlein
,
M.
,
Price
,
S.
,
Ahmad
,
F.
,
Gulari
,
E.
,
Tiedje
,
J. M.
, and
Hashsham
,
S. A.
, 2012
, “
Gene-Z: A Device for Point of Care Genetic Testing Using a Smartphone
,” Lab. Chip
,
12
(8
), pp. 1454
–1462
.10.1039/c2lc21226a48.
Velders
,
A. H.
,
Schoen
,
C.
, and
Saggiomo
,
V.
, 2018
, “
Loop-Mediated Isothermal Amplification (LAMP) Shield for Arduino DNA Detection
,” BMC Res. Notes
,
11
(1
), p. 93
.10.1186/s13104-018-3197-949.
Schultz
,
A. L.
, 2021
, A Portable Point-of-Care Device Using Joule Heating and Latent Energy Storage for the Temperature Regulation of Isothermal Nucleic Acid Amplification Tests
,
University of Arkansas
, Fayetteville, AR.
