Rapid detection of Mycobacterium tuberculosis cells by using microtip-based immunoassay

Anal Bioanal Chem. 2009 Mar;393(6-7):1593-600. doi: 10.1007/s00216-008-2591-x. Epub 2009 Jan 13.

Abstract

This paper describes a microtip-based approach of concentrating target analytes for a highly sensitive bioassay. As an example, rapid screening of bacterial whole cells is presented to detect Mycobacterium tuberculosis (MTB), a pathogenic bacterium for human tuberculosis (TB). The concentration and detection is performed with three sequential steps of (1) attracting bacterial whole cells in the vicinity of a microtip by alternating current electroosmotic flow; (2) capturing the cells onto the microtip by capillary action; (3) binding fluorophore-labeled polyclonal antibodies to the cells followed by fluorescence measurement (immunofluorescence). Through this mechanism, MTB cells have been detected to the concentration of 8,000 cells/mL within 10 min. This sensitivity is comparable to that of Ziehl-Neelsen smear microscopy, a common culture-free screening method for diagnosis of TB. For comparison, Escherichia coli O157:H7 cells have also been detected to the concentration of 30,000 cells/mL in the same way.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Fluorescent Antibody Technique / methods
  • Immunoassay / methods*
  • Microchip Analytical Procedures / methods*
  • Microelectrodes
  • Mycobacterium tuberculosis / cytology*
  • Sensitivity and Specificity