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.
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 entericaTyphi, 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.
Authors: Steffen Zehnle; Frank Schwemmer; Günter Roth; Felix von Stetten; Roland Zengerle; Nils Paust Journal: Lab Chip Date: 2012-12-21 Impact factor: 6.799
Authors: Shuenn-Jue Wu; Subhamoy Pal; Sajeewane Ekanayake; David Greenwald; Silvia Lara; Kanakatte Raviprakash; Tadeusz Kochel; Kevin Porter; Curtis Hayes; William Nelson; Johnny Callahan Journal: Am J Trop Med Hyg Date: 2008-10 Impact factor: 2.345
Authors: Herman Kosasih; Susana Widjaja; Edwin Surya; Sri H Hadiwijaya; Deni P R Butarbutar; Ungke A Jaya; Bachti Alisjahbana; Maya Williams Journal: Southeast Asian J Trop Med Public Health Date: 2012-01 Impact factor: 0.267
Authors: Claudia Rückert; James Weger-Lucarelli; Selene M Garcia-Luna; Michael C Young; Alex D Byas; Reyes A Murrieta; Joseph R Fauver; Gregory D Ebel Journal: Nat Commun Date: 2017-05-19 Impact factor: 14.919
Authors: Konstantinos Mitsakakis; Sebastian Hin; Pie Müller; Nadja Wipf; Edward Thomsen; Michael Coleman; Roland Zengerle; John Vontas; Konstantinos Mavridis Journal: Int J Environ Res Public Health Date: 2018-02-03 Impact factor: 3.390
Authors: John A Crump; Anne B Morrissey; William L Nicholson; Robert F Massung; Robyn A Stoddard; Renee L Galloway; Eng Eong Ooi; Venance P Maro; Wilbrod Saganda; Grace D Kinabo; Charles Muiruri; John A Bartlett Journal: PLoS Negl Trop Dis Date: 2013-07-18
Authors: Yukari C Manabe; Joshua Betz; Olivia Jackson; Victor Asoala; Isabel Bazan; Paul W Blair; Aileen Chang; Sarunyou Chusri; John A Crump; Kimberly A Edgel; Dennis J Faix; Stefan Fernandez; Anne T Fox; Jose A Garcia; Max Grogl; Erin A Hansen; Vireak Heang; Stacey L House; Krisada Jongsakul; Michael B Kaburise; Chonticha Klungthong; Mohammed Lamorde; Andrew G Letizia; Ivette Lorenzana; Malen Luy; Vanance P Maro; Christopher N Mores; Christopher A Myers; Abraham R Oduro; Leda Parham; Abigail J Porzucek; Michael Prouty; David S Rabiger; Matthew P Rubach; Crystyan Siles; Maria Silva; Chinaka Ukachu; John N Waitumbi; Cynthia L Phillips; Brian W Jones Journal: Lancet Infect Dis Date: 2022-06-15 Impact factor: 71.421
Authors: Desirée Baumgartner; Benita Johannsen; Mara Specht; Jan Lüddecke; Markus Rombach; Sebastian Hin; Nils Paust; Felix von Stetten; Roland Zengerle; Christopher Herz; Johannes R Peham; Pune N Paqué; Thomas Attin; Joël S Jenzer; Philipp Körner; Patrick R Schmidlin; Thomas Thurnheer; Florian J Wegehaupt; Wendy E Kaman; Andrew Stubbs; John P Hays; Viorel Rusu; Alex Michie; Thomas Binsl; David Stejskal; Michal Karpíšek; Kai Bao; Nagihan Bostanci; Georgios N Belibasakis; Konstantinos Mitsakakis Journal: Biosensors (Basel) Date: 2021-10-28