Advertisement

Use of Immunomagnetic Carriers for the Rapid Detection of Virulent Bacteria from Biological Samples

Chapter

Abstract

An antibody-based magnetic immunoassay for the effective and rapid detection of virulent bacteria from biological samples was developed. Chemically linked streptavidin-magnetic particles and biotinylated primary antibodies as immunomagnetic carriers were used for bacteria capture. A probe-labeled secondary antibody formed a sandwich for antigen detection. The use of immunomagnetic carriers for bacteria separation is a key step to reduce media interference from biological samples prior to detection. Magnetic separation has been achieved by two types of separators developed in-house. The first is a single channeled flow-through immunomagnetic separator (IMS), the second a multiple-well magnetic plate (MP) separator. An electrochemiluminescence analyzer and a fluorescence plate reader were employed to detect the bacteria. Experiments were performed for virulent Escherichia coli 0157:H7 and Salmonella typhimurium strains, Bacillus anthracis and Bacillus subtilus var. niger (BG) spore detection from different biological samples using both magnetic separators. In general, the detection of the biological samples after magnetic separation was more sensitive and showed lower background levels than the detection without magnetic separation.

Keywords

Magnetic Particle Magnetic Separation Fluorescence Plate Reader Bacterium Detection Virulent Bacterium 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Lamothe F, Gaudreau C, Bernard D, and Gill S (1983). Hemorrhagic colitis following the consumption of hamburgers. Quebec. Can. Dis. Weekly Rep. 9, 50–51.Google Scholar
  2. 2.
    LaForce FM (1978). Woolsorters’disease in England. Bull. NY Acad. Med. 54 956.Google Scholar
  3. 3.
    Hill WE, and Carlisle CL (1981). Loss of plasmids during enrichment for Escherichia coli. Applied Environmental Microbiology, 41 1046–1048.Google Scholar
  4. 4.
    Lo YM, Mehal WZ and Fleming KA (1988). False-positive results and the polymerase chain reaction. Lancet II, 679.Google Scholar
  5. 5.
    Olsvik O, Popovic T, Skjerve E, Cudjoe KS, Homes E, Ugelstad J, and Uhlen M (1994). Magnetic separation techniques in diagnostic microbiology. Clinical Microbiology Reviews 7, 43–54.Google Scholar
  6. 6.
    Cudjoe KS, Thorsen LI, Sorensen T and Reseland J (1991). Detection of Clostridium perfringens types A enterotoxin in faecal and food samples using immunomagnetic separation (1MS)-ELISA. Int. J. Food Microbiol. 12, 313–322.Google Scholar
  7. 7.
    Yu H and Bruno JB (1996). Immunomagnetic electrochemiluminescent detection of Escherichia coli 0157 and Salmonella typhimurium in foods and environmental water samples. Applied Environmental Microbiology. 62 587–592.Google Scholar
  8. 8.
    Yu H and Bruno JG (1995). Detection of the coliforrn bacteria Escherichia coli and Salmonella sp. in water by a sensitive and rapid immunomagnetic electrochemiluminescence (ECL) technique. EUROPT Series for Environmental Monitoring and Hazardous Waste Site remediation, SPIE Vol. 2504, 240–252.Google Scholar
  9. 9.
    Yu H (1996). Enhancing immuno-electrochemiluminescence for sensitive bacterial detection. Journal of Immunological Methods, 192 63–71.Google Scholar
  10. 10.
    Cano RJ, Torres MI, Klem RE and Palomares JC (1992). DNA hybridization assay using AttoPhos, afluorescent substrate for alkaline phophatase. Biotechniques. 12, 264–268.Google Scholar

Copyright information

© Springer Science+Business Media New York 1997

Authors and Affiliations

  • Hao Yu
    • 1
  1. 1.Edgewood Research, Development, and Engineering Center, Aberdeen Proving GroundCalspan SRL Corp.USA

Personalised recommendations