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Integrated Acoustic Separation, Enrichment, and Microchip Polymerase Chain Reaction Detection of Bacteria from Blood for Rapid Sepsis Diagnostics

Ohlsson, Pelle LU ; Evander, Mikael LU ; Petersson, Klara LU ; Mellhammar, Lisa LU ; Lehmusvuori, Ari; Karhunen, Ulla; Soikkeli, Minna; Seppä, Titta; Tuunainen, Emilia and Spangar, Anni, et al. (2016) In Analytical Chemistry 88(19). p.9403-9411
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 are trapped onto suspended polystyrene particles. The trapped bacteria are subsequently washed while held in the acoustic trap and released into a polymer microchip containing dried polymerase chain reaction (PCR) reagents, followed by thermocycling for... (More)

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 are trapped onto suspended polystyrene particles. The trapped bacteria are subsequently 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 a further developed acoustic sample preparation system may open the door for a new and faster automated method to diagnose sepsis.

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published
subject
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Analytical Chemistry
volume
88
issue
19
pages
9 pages
publisher
The American Chemical Society
external identifiers
  • scopus:84989814539
  • wos:000384842200014
ISSN
0003-2700
DOI
10.1021/acs.analchem.6b00323
language
English
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yes
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3ea183af-5574-43c3-9dfc-7932efd4e37b
date added to LUP
2016-10-21 08:16:51
date last changed
2017-10-29 04:53:02
@article{3ea183af-5574-43c3-9dfc-7932efd4e37b,
  abstract     = {<p>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 are trapped onto suspended polystyrene particles. The trapped bacteria are subsequently 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 a further developed acoustic sample preparation system may open the door for a new and faster automated method to diagnose sepsis.</p>},
  author       = {Ohlsson, Pelle and Evander, Mikael and Petersson, Klara and Mellhammar, Lisa and Lehmusvuori, Ari and Karhunen, Ulla and Soikkeli, Minna and Seppä, Titta and Tuunainen, Emilia and Spangar, Anni and Von Lode, Piia and Rantakokko-Jalava, Kaisu and Otto, Gisela and Scheding, Stefan and Soukka, Tero and Wittfooth, Saara and Laurell, Thomas},
  issn         = {0003-2700},
  language     = {eng},
  month        = {10},
  number       = {19},
  pages        = {9403--9411},
  publisher    = {The American Chemical Society},
  series       = {Analytical Chemistry},
  title        = {Integrated Acoustic Separation, Enrichment, and Microchip Polymerase Chain Reaction Detection of Bacteria from Blood for Rapid Sepsis Diagnostics},
  url          = {http://dx.doi.org/10.1021/acs.analchem.6b00323},
  volume       = {88},
  year         = {2016},
}