A microfluidic platform combined with bacteriophage receptor binding proteins for multiplex detection of Escherichia coli and Pseudomonas aeruginosa in blood

Costa, Susana P.; Caneira, Catarina R.F.; Chu, V.; Freitas, P. P.; Conde, J.P.; Carvalho, Cristiana

doi:10.1016/j.snb.2022.132917

Cited by 10 publications

(3 citation statements)

References 92 publications

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“…We have previously demonstrated the high specificity of RBPs for other nosocomial pathogens such as Enterococcus , S . aureus , Pseudomonas aeruginosa , and Klebsiella pneumoniae [ 21 , 29 , 30 ]. Moreover, Gp17 has been already tested in different human specimens enabling the specific detection of E .…”

Section: Resultsmentioning

confidence: 99%

“…This is of utmost importance since the complexity of blood frequently hampers the efficiency of several methodologies currently used for the detection of pathogens. Examples include the interference of blood components in molecular methods and the autofluorescence of erythrocytes that affect optical-based diagnostic techniques [ 12 , 21 ].…”

Section: Resultsmentioning

confidence: 99%

“…As the phages normally infect only one species of host bacteria, the RBPs present high binding specificity and offer low production cost, facility for genetic modification, and high chemical and physical resistance [ 19 ]. Although the effective use of these proteins as affinity molecules has been reported for different pathogens [ 19 , 20 ], to the best of our knowledge, their application in microfluidic platforms for human specimen analysis has only been described by our group [ 21 ]. Overall, the requirement still exists for a cost-effective, fast, selective, and reliable platform to timely respond to the needs of targeting BSIs, especially, in intensive care units (ICUs).…”

Section: Introductionmentioning

confidence: 99%

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A magnetic nanoparticle-based microfluidic device fabricated using a 3D-printed mould for separation of Escherichia coli from blood

Jóskowiak,

Nogueira,

Costa

et al. 2023

Microchim Acta

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Herein, A microfluidic device is described, produced with a 3D-printed master mould that rapidly separates and concentrates Escherichia coli directly from whole blood samples, enabling a reduction in the turnaround time of bloodstream infections (BSIs) diagnosis. Moreover, it promotes the cleansing of the blood samples whose complexity frequently hampers bacterial detection. The device comprises a serpentine mixing channel with two inlets, one for blood samples (spiked with bacteria) and the other for magnetic nanoparticles (MNPs) functionalized with a (bacterio)phage receptor-binding protein (RBP) with high specificity for E. coli. After the magnetic labelling of bacteria throughout the serpentine, the microchannel ends with a trapping reservoir where bacteria-MNPs conjugates are concentrated using a permanent magnet. The optimized sample preparation device successfully recovered E. coli (on average, 66%) from tenfold diluted blood spiked within a wide range of bacterial load (102 CFU to 107 CFU mL−1). The non-specific trapping, tested with Staphylococcus aureus, was at a negligible level of 12%. The assay was performed in 30 min directly from diluted blood thus presenting an advantage over the conventional enrichment in blood cultures (BCs). The device is simple and cheap to fabricate and can be tailored for multiple bacterial separation from complex clinical samples by using RBPs targeting different species. Moreover, the possibility to integrate a biosensing element to detect bacteria on-site can provide a reliable, fast, and cost-effective point-of-care device. Graphical abstract

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%