A1 Refereed original research article in a scientific journal
Integrated Acoustic Separation, Enrichment, and Microchip Polymerase Chain Reaction Detection of Bacteria from Blood for Rapid Sepsis Diagnostics
Authors: Ohlsson P, Evander M, Petersson K, Mellhammar L, Lehmusvuori A, Karhunen U, Soikkeli M, Seppa T, Tuunainen E, Spangar A, von Lode P, Rantakokko-Jalava K, Otto G, Scheding S, Soukka T, Wittfooth S, Laurell T
Publisher: AMER CHEMICAL SOC
Publication year: 2016
Journal: Analytical Chemistry
Journal name in source: ANALYTICAL CHEMISTRY
Journal acronym: ANAL CHEM
Volume: 88
Issue: 19
First page : 9403
Last page: 9411
Number of pages: 9
ISSN: 0003-2700
DOI: https://doi.org/10.1021/acs.analchem.6b00323
Abstract
This paper describes an integrated,microsystem for rapid separation, enrichment, and detection of bacteria from blood, addressing the unmet clinical need for rapid sepsis diagnostics. The blood sample is first processed in an acoustophoresis chip, where red blood cells are focused to the center of the channel by an acoustic standing wave and sequentially removed. The, bacteria-containing plasma proceeds to a glass capillary with a localized acoustic standing wave field where the bacteria ate trapped onto suspended polystyrene particles. The trapped bacteria are subequently washed while held in the acoustic trap and released into a polymer microchip containing dried polymerase chain reaction (PCR) reagents, followed by thermocycling for target sequence, amplification. The entire process is completed in less than 2 h. Testing with Pseudomonas putida spiked into whole blood revealed a detection limit of 1000 bacteria/mL for this first-generation analysis system. In samples from septic patients,. the system was able to detect Escherichia coli in half of the cases identified by blood culture. This indicates that the current system detects bacteria in patient,samples in the upper part of the of clinically relevant bacteria concentration range and that further developed acoustic sample preparation system may open the door for a new and faster automated method to diagnose sepsis.
This paper describes an integrated,microsystem for rapid separation, enrichment, and detection of bacteria from blood, addressing the unmet clinical need for rapid sepsis diagnostics. The blood sample is first processed in an acoustophoresis chip, where red blood cells are focused to the center of the channel by an acoustic standing wave and sequentially removed. The, bacteria-containing plasma proceeds to a glass capillary with a localized acoustic standing wave field where the bacteria ate trapped onto suspended polystyrene particles. The trapped bacteria are subequently washed while held in the acoustic trap and released into a polymer microchip containing dried polymerase chain reaction (PCR) reagents, followed by thermocycling for target sequence, amplification. The entire process is completed in less than 2 h. Testing with Pseudomonas putida spiked into whole blood revealed a detection limit of 1000 bacteria/mL for this first-generation analysis system. In samples from septic patients,. the system was able to detect Escherichia coli in half of the cases identified by blood culture. This indicates that the current system detects bacteria in patient,samples in the upper part of the of clinically relevant bacteria concentration range and that further developed acoustic sample preparation system may open the door for a new and faster automated method to diagnose sepsis.
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