Immunodetection of inactivated Francisella tularensis bacteria by using a quartz crystal microbalance with dissipation monitoring

Anal Bioanal Chem. 2012 Aug;404(3):843-51. doi: 10.1007/s00216-012-6172-7. Epub 2012 Jun 22.

Abstract

Francisella tularensis are very small, gram-negative bacteria which are capable of infecting a number of mammals. As a highly pathogenic species, it is a potential bioterrorism agent. In this work we demonstrate a fast immunological detection system for whole F. tularensis bacteria. The technique is based on a quartz crystal microbalance with dissipation monitoring (QCMD), which uses sensor chips modified by a specific antibody. This antibody is useful as a capture molecule to capture the lipopolysaccharide structure on the surface of the bacterial cell wall. The QCMD technique is combined with a microfluidic system and allows the label-free online detection of the binding of whole bacteria to the sensor surface in a wide dynamic concentration range. A detection limit of about 4 × 10(3) colony-forming units per milliliter can be obtained. Furthermore, a rather short analysis time and a clear discrimination against other bacteria can be achieved. Additionally, we demonstrate two possibilities for specific and significant signal enhancement by using antibody-functionalized gold nanoparticles or an enzymatic precipitation reaction. These additional steps can be seen as further proof of the specificity and validity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Immobilized / chemistry*
  • Antibodies, Immobilized / immunology
  • Biosensing Techniques*
  • Francisella tularensis / chemistry
  • Francisella tularensis / immunology
  • Francisella tularensis / isolation & purification*
  • Gold / chemistry
  • Limit of Detection
  • Lipopolysaccharides / chemistry*
  • Metal Nanoparticles / chemistry
  • Microfluidic Analytical Techniques
  • Quartz Crystal Microbalance Techniques / instrumentation
  • Quartz Crystal Microbalance Techniques / methods*
  • Stem Cells

Substances

  • Antibodies, Immobilized
  • Lipopolysaccharides
  • Gold