Format

Send to

Choose Destination
See comment in PubMed Commons below
PLoS One. 2014 Jan 22;9(1):e86018. doi: 10.1371/journal.pone.0086018. eCollection 2014.

Semi-automated, occupationally safe immunofluorescence microtip sensor for rapid detection of Mycobacterium cells in sputum.

Author information

1
Department of Mechanical Engineering, University of Washington, Seattle, Washington, United States of America.
2
Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, Washington, United States of America.
3
Departments of Medicine-Division of Medical Genetics and Genome Sciences, University of Washington, Seattle, Washington, United States of America.
4
NanoFacture, Inc., Bellevue, Washington, United States of America.

Abstract

An occupationally safe (biosafe) sputum liquefaction protocol was developed for use with a semi-automated antibody-based microtip immunofluorescence sensor. The protocol effectively liquefied sputum and inactivated microorganisms including Mycobacterium tuberculosis, while preserving the antibody-binding activity of Mycobacterium cell surface antigens. Sputum was treated with a synergistic chemical-thermal protocol that included moderate concentrations of NaOH and detergent at 60°C for 5 to 10 min. Samples spiked with M. tuberculosis complex cells showed approximately 10(6)-fold inactivation of the pathogen after treatment. Antibody binding was retained post-treatment, as determined by analysis with a microtip immunosensor. The sensor correctly distinguished between Mycobacterium species and other cell types naturally present in biosafe-treated sputum, with a detection limit of 100 CFU/mL for M. tuberculosis, in a 30-minute sample-to-result process. The microtip device was also semi-automated and shown to be compatible with low-cost, LED-powered fluorescence microscopy. The device and biosafe sputum liquefaction method opens the door to rapid detection of tuberculosis in settings with limited laboratory infrastructure.

PMID:
24465845
PMCID:
PMC3899086
DOI:
10.1371/journal.pone.0086018
[Indexed for MEDLINE]
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Public Library of Science Icon for PubMed Central
    Loading ...
    Support Center