Review on biphasic blood drying method for rapid pathogen detection in bloodstream infections

Rapid and accurate detection of pathogenic microorganisms in blood is critical for diagnosing life-threatening conditions such as bloodstream infections (BSIs). Current methods for the detection and identification of bacteria from large volumes of blood (5 mL) involve culture steps followed by DNA e...

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Main Authors: Jongwon Lim, Katherine Koprowski, Matthew Wester, Enrique Valera, Rashid Bashir
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:SLAS Technology
Online Access:http://www.sciencedirect.com/science/article/pii/S2472630325000342
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author Jongwon Lim
Katherine Koprowski
Matthew Wester
Enrique Valera
Rashid Bashir
author_facet Jongwon Lim
Katherine Koprowski
Matthew Wester
Enrique Valera
Rashid Bashir
author_sort Jongwon Lim
collection DOAJ
description Rapid and accurate detection of pathogenic microorganisms in blood is critical for diagnosing life-threatening conditions such as bloodstream infections (BSIs). Current methods for the detection and identification of bacteria from large volumes of blood (5 mL) involve culture steps followed by DNA extraction/purification/concentration and Polymerase Chain Reaction (PCR)-based nucleic acid amplification. DNA extraction and amplification directly from blood samples is hampered by the complexity of the blood matrix, resulting in time-consuming and labor-intensive processes. This review delves into recent advancements in molecular diagnostics based on blood drying, coined as ‘biphasic reaction’, and highlights this new technique that attempts to overcome the limitations of traditional sample preparation and amplification processes. The biphasic blood drying method, in combination with isothermal amplification methods such as loop-mediated isothermal amplification (LAMP) or recombinase polymerase amplification (RPA), has recently been shown to improve the sensitivity of detection of bacterial, viral, and fungal pathogens from ∼1 mL of whole blood, while minimizing DNA loss and avoiding the use of extraction/purification/concentration kits. Furthermore, the biphasic approach in combination with LAMP has been shown to be a culture-free method capable of detecting bacteria in clinical samples with a sensitivity of ∼1 CFU/mL in ∼2.5 h. This represents a significant reduction in detection and identification time compared to current clinical procedures based on bacterial culture prior to PCR amplification. This review paper aims to be a guide to identify new opportunities for future advancements and applications of the biphasic technology.
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spelling doaj-art-e1f8b512e387418db1ce2b9b046078d82025-08-20T03:42:15ZengElsevierSLAS Technology2472-63032025-06-013210027610.1016/j.slast.2025.100276Review on biphasic blood drying method for rapid pathogen detection in bloodstream infectionsJongwon Lim0Katherine Koprowski1Matthew Wester2Enrique Valera3Rashid Bashir4Department of Bioengineering, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USA; Nick Holonyak Jr. Micro and Nanotechnology Laboratory, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USA; Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USA; Material Research Laboratory, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USADepartment of Bioengineering, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USA; Nick Holonyak Jr. Micro and Nanotechnology Laboratory, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USADepartment of Bioengineering, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USA; Nick Holonyak Jr. Micro and Nanotechnology Laboratory, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USADepartment of Bioengineering, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USA; Nick Holonyak Jr. Micro and Nanotechnology Laboratory, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USA; Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USA; Corresponding authors.Department of Bioengineering, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USA; Nick Holonyak Jr. Micro and Nanotechnology Laboratory, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USA; Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USA; Department of Materials Science and Engineering, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USA; Department of Electrical and Computer Engineering, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USA; Department of Mechanical Science and Engineering, University of Illinois at Urbana–Champaign, Urbana, IL 61801, USA; Department of Biomedical and Translational Sciences, Carle Illinois College of Medicine, Urbana, IL 61801, USA; Chan Zuckerberg Biohub Chicago, Chicago, IL 60642, USA.; Corresponding authors.Rapid and accurate detection of pathogenic microorganisms in blood is critical for diagnosing life-threatening conditions such as bloodstream infections (BSIs). Current methods for the detection and identification of bacteria from large volumes of blood (5 mL) involve culture steps followed by DNA extraction/purification/concentration and Polymerase Chain Reaction (PCR)-based nucleic acid amplification. DNA extraction and amplification directly from blood samples is hampered by the complexity of the blood matrix, resulting in time-consuming and labor-intensive processes. This review delves into recent advancements in molecular diagnostics based on blood drying, coined as ‘biphasic reaction’, and highlights this new technique that attempts to overcome the limitations of traditional sample preparation and amplification processes. The biphasic blood drying method, in combination with isothermal amplification methods such as loop-mediated isothermal amplification (LAMP) or recombinase polymerase amplification (RPA), has recently been shown to improve the sensitivity of detection of bacterial, viral, and fungal pathogens from ∼1 mL of whole blood, while minimizing DNA loss and avoiding the use of extraction/purification/concentration kits. Furthermore, the biphasic approach in combination with LAMP has been shown to be a culture-free method capable of detecting bacteria in clinical samples with a sensitivity of ∼1 CFU/mL in ∼2.5 h. This represents a significant reduction in detection and identification time compared to current clinical procedures based on bacterial culture prior to PCR amplification. This review paper aims to be a guide to identify new opportunities for future advancements and applications of the biphasic technology.http://www.sciencedirect.com/science/article/pii/S2472630325000342
spellingShingle Jongwon Lim
Katherine Koprowski
Matthew Wester
Enrique Valera
Rashid Bashir
Review on biphasic blood drying method for rapid pathogen detection in bloodstream infections
SLAS Technology
title Review on biphasic blood drying method for rapid pathogen detection in bloodstream infections
title_full Review on biphasic blood drying method for rapid pathogen detection in bloodstream infections
title_fullStr Review on biphasic blood drying method for rapid pathogen detection in bloodstream infections
title_full_unstemmed Review on biphasic blood drying method for rapid pathogen detection in bloodstream infections
title_short Review on biphasic blood drying method for rapid pathogen detection in bloodstream infections
title_sort review on biphasic blood drying method for rapid pathogen detection in bloodstream infections
url http://www.sciencedirect.com/science/article/pii/S2472630325000342
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