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Microfluidic-Based Bacteria Isolation from Whole Blood for Diagnostics of Blood Stream Infection

  • Sergey Zelenin
  • Harisha Ramachandraiah
  • Asim Faridi
  • Aman RussomEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1547)

Abstract

Bacterial blood stream infection (BSI) potentially leads to life-threatening clinical conditions and medical emergencies such as severe sepsis, septic shock, and multi organ failure syndrome. Blood culturing is currently the gold standard for the identification of microorganisms and, although it has been automated over the decade, the process still requires 24–72 h to complete. This long turnaround time, especially for the identification of antimicrobial resistance, is driving the development of rapid molecular diagnostic methods. Rapid detection of microbial pathogens in blood related to bloodstream infections will allow the clinician to decide on or adjust the antimicrobial therapy potentially reducing the morbidity, mortality, and economic burden associated with BSI. For molecular-based methods, there is a lot to gain from an improved and straightforward method for isolation of bacteria from whole blood for downstream processing.

We describe a microfluidic-based sample-preparation approach that rapidly and selectively lyses all blood cells while it extracts intact bacteria for downstream analysis. Whole blood is exposed to a mild detergent, which lyses most blood cells, and then to osmotic shock using deionized water, which eliminates the remaining white blood cells. The recovered bacteria are 100 % viable, which opens up possibilities for performing drug susceptibility tests and for nucleic-acid-based molecular identification.

Key words

Sepsis Blood stream infection Microfluidics Diagnostic Pathogens Bacteremia Point-of-care diagnostics 

Notes

Acknowledgment

This work was supported by the European Commission (projects FP7 InTopSens and IMI RAPP-ID)

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Sergey Zelenin
    • 1
  • Harisha Ramachandraiah
    • 1
  • Asim Faridi
    • 1
  • Aman Russom
    • 1
    Email author
  1. 1.Science for Life Laboratory, Division of Proteomics and NanobiotechnologyKTH Royal Institute of TechnologySolnaSweden

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