Fully automated point-of-care differential diagnosis of acute febrile illness.
Journal
PLoS neglected tropical diseases
ISSN: 1935-2735
Titre abrégé: PLoS Negl Trop Dis
Pays: United States
ID NLM: 101291488
Informations de publication
Date de publication:
02 2021
02 2021
Historique:
received:
06
01
2020
accepted:
26
01
2021
entrez:
25
2
2021
pubmed:
26
2
2021
medline:
23
6
2021
Statut:
epublish
Résumé
In this work, a platform was developed and tested to allow to detect a variety of candidate viral, bacterial and parasitic pathogens, for acute fever of unknown origin. The platform is based on a centrifugal microfluidic cartridge, the LabDisk ("FeverDisk" for the specific application), which integrates all necessary reagents for sample-to-answer analysis and is processed by a compact, point-of-care compatible device. A sample volume of 200 μL per FeverDisk was used. In situ extraction with pre-stored reagents was achieved by bind-wash-elute chemistry and magnetic particles. Enzymes for the loop-mediated isothermal amplification (LAMP) were pre-stored in lyopellet form providing stability and independence from the cold chain. The total time to result from sample inlet to read out was 2 h. The proof-of-principle was demonstrated in three small-scale feasibility studies: in Dakar, Senegal and Khartoum, Sudan we tested biobanked samples using 29 and 9 disks, respectively; in Reinfeld, Germany we tested spiked samples and analyzed the limit of detection using three bacteria simultaneously spiked in whole blood using 15 disks. Overall during the three studies, the FeverDisk detected dengue virus (different serotypes), chikungunya virus, Plasmodium falciparum, Salmonella enterica Typhi, Salmonella enterica Paratyphi A and Streptococcus pneumoniae. The FeverDisk proved to be universally applicable as it successfully detected all different types of pathogens as single or co-infections, while it also managed to define the serotype of un-serotyped dengue samples. Thirty-eight FeverDisks at the two African sites provided 59 assay results, out of which 51 (86.4%) were confirmed with reference assay results. The results provide a promising outlook for future implementation of the platform in larger prospective clinical studies for defining its clinical sensitivity and specificity. The technology aims to provide multi-target diagnosis of the origins of fever, which will help fight lethal diseases and the incessant rise of antimicrobial resistance.
Sections du résumé
BACKGROUND
In this work, a platform was developed and tested to allow to detect a variety of candidate viral, bacterial and parasitic pathogens, for acute fever of unknown origin. The platform is based on a centrifugal microfluidic cartridge, the LabDisk ("FeverDisk" for the specific application), which integrates all necessary reagents for sample-to-answer analysis and is processed by a compact, point-of-care compatible device.
METHODOLOGY/PRINCIPAL FINDINGS
A sample volume of 200 μL per FeverDisk was used. In situ extraction with pre-stored reagents was achieved by bind-wash-elute chemistry and magnetic particles. Enzymes for the loop-mediated isothermal amplification (LAMP) were pre-stored in lyopellet form providing stability and independence from the cold chain. The total time to result from sample inlet to read out was 2 h. The proof-of-principle was demonstrated in three small-scale feasibility studies: in Dakar, Senegal and Khartoum, Sudan we tested biobanked samples using 29 and 9 disks, respectively; in Reinfeld, Germany we tested spiked samples and analyzed the limit of detection using three bacteria simultaneously spiked in whole blood using 15 disks. Overall during the three studies, the FeverDisk detected dengue virus (different serotypes), chikungunya virus, Plasmodium falciparum, Salmonella enterica Typhi, Salmonella enterica Paratyphi A and Streptococcus pneumoniae.
CONCLUSIONS/SIGNIFICANCE
The FeverDisk proved to be universally applicable as it successfully detected all different types of pathogens as single or co-infections, while it also managed to define the serotype of un-serotyped dengue samples. Thirty-eight FeverDisks at the two African sites provided 59 assay results, out of which 51 (86.4%) were confirmed with reference assay results. The results provide a promising outlook for future implementation of the platform in larger prospective clinical studies for defining its clinical sensitivity and specificity. The technology aims to provide multi-target diagnosis of the origins of fever, which will help fight lethal diseases and the incessant rise of antimicrobial resistance.
Identifiants
pubmed: 33630852
doi: 10.1371/journal.pntd.0009177
pii: PNTD-D-19-02168
pmc: PMC7906357
doi:
Substances chimiques
DNA, Bacterial
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0009177Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
Références
PLoS Negl Trop Dis. 2013 Jul 18;7(7):e2324
pubmed: 23875053
PLoS Negl Trop Dis. 2014 Mar 06;8(3):e2730
pubmed: 24603874
Nat Commun. 2017 May 19;8:15412
pubmed: 28524874
Lab Chip. 2010 Jul 21;10(14):1758-73
pubmed: 20512178
Lab Chip. 2012 Dec 21;12(24):5142-5
pubmed: 23108455
BMC Res Notes. 2018 Feb 1;11(1):93
pubmed: 29391052
Int J Environ Res Public Health. 2018 Feb 03;15(2):
pubmed: 29401670
Southeast Asian J Trop Med Public Health. 2012 Jan;43(1):55-61
pubmed: 23082554
Nat Protoc. 2008;3(5):877-82
pubmed: 18451795
Am J Trop Med Hyg. 2008 Oct;79(4):505-10
pubmed: 18840736
Crit Rev Clin Lab Sci. 2016;53(2):87-105
pubmed: 26376713
PLoS Negl Trop Dis. 2013 Jul 25;7(7):e2331
pubmed: 23936565
PLoS One. 2017 Dec 21;12(12):e0189968
pubmed: 29267339
Lab Chip. 2018 Jan 16;18(2):362-370
pubmed: 29297912
Malar J. 2015 Jul 19;14:281
pubmed: 26186936
PLoS One. 2016 Jan 28;11(1):e0146054
pubmed: 26821179
Clin Infect Dis. 2006 Apr 15;42(8):1127-34
pubmed: 16575730
Lab Chip. 2017 Feb 28;17(5):864-875
pubmed: 28181607
PLoS One. 2012;7(2):e31432
pubmed: 22384022
PLoS One. 2014 Aug 04;9(8):e104197
pubmed: 25090651
Korean J Parasitol. 2009 Jun;47(2):93-102
pubmed: 19488414
Travel Med Infect Dis. 2016 May-Jun;14(3):177-9
pubmed: 27224471
Trends Microbiol. 2014 Mar;22(3):138-46
pubmed: 24468533
Methods Mol Biol. 2020;2142:147-164
pubmed: 32367366
BMC Bioinformatics. 2011 Jun 16;12:240
pubmed: 21679460
Lancet Glob Health. 2013 Jul;1(1):e46-54
pubmed: 24748368
Nat Rev Microbiol. 2006 Sep;4(9 Suppl):S34-8
pubmed: 17034070
Cochrane Database Syst Rev. 2017 May 26;5:CD008892
pubmed: 28545155
Clin Microbiol Infect. 2013 May;19(5):422-31
pubmed: 23413992
BMC Infect Dis. 2018 Jan 30;18(1):61
pubmed: 29382300
J Pharm Bioallied Sci. 2013 Oct;5(4):245-52
pubmed: 24302831
Lab Chip. 2016 Jan 7;16(1):199-207
pubmed: 26610171
PLoS Negl Trop Dis. 2018 May 29;12(5):e0006448
pubmed: 29813065
Lab Chip. 2015;15(18):3749-59
pubmed: 26235430
J Virol Methods. 2005 Mar;124(1-2):65-71
pubmed: 15664052
BMJ. 2004 Nov 20;329(7476):1212
pubmed: 15542534
Biochem Biophys Res Commun. 2001 Nov 23;289(1):150-4
pubmed: 11708792
PLoS Negl Trop Dis. 2014 Oct 16;8(10):e3171
pubmed: 25330157
J Clin Microbiol. 1990 Mar;28(3):495-503
pubmed: 1691208
N Engl J Med. 2014 Feb 27;370(9):809-17
pubmed: 24571753
Pharmacogenomics. 2013 Jul;14(9):1011-5
pubmed: 23837475
Clin Infect Dis. 2012 Apr;54(8):1107-9
pubmed: 22357701
PLoS Negl Trop Dis. 2014 Jul 31;8(7):e3043
pubmed: 25078474
Lab Chip. 2013 Aug 7;13(15):2888-92
pubmed: 23674222
Microelectron Eng. 2018 Dec 5;201:26-59
pubmed: 32287568
PLoS Negl Trop Dis. 2018 May 29;12(5):e0006381
pubmed: 29813062
J Infect Dis. 2014 Oct 15;210(8):1180-7
pubmed: 24795480
Stud Health Technol Inform. 2016;224:54-60
pubmed: 27225553
Chem Soc Rev. 2015 Oct 7;44(17):6187-229
pubmed: 26035697