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Comparing recovering efficiency of immunomagnetic separation and centrifugation of mycobacteria in metalworking fluids

  • Environmental Biotechnology
  • Published:
Journal of Industrial Microbiology and Biotechnology

Abstract

The accurate detection and enumeration of Mycobacterium immunogenum in metalworking fluids (MWFs) is imperative from an occupational health and industrial fluids management perspective. We report here a comparison of immunomagnetic separation (IMS) coupled to flow-cytometric enumeration, with traditional centrifugation techniques for mycobacteria in a semisynthetic MWF. This immunolabeling involves the coating of laboratory-synthesized nanometer-scale magnetic particles with protein A, to conjugate a primary antibody (Ab), specific to Mycobacterium spp. By using magnetic separation and flow-cytometric quantification, this approach enabled much higher recovery efficiency and fluorescent light intensities in comparison to the widely applied centrifugation technique. This IMS technique increased the cell recovery efficiency by one order of magnitude, and improved the fluorescence intensity of the secondary Ab conjugate by 2-fold, as compared with traditional techniques. By employing nanometer-scale magnetic particles, IMS was found to be compatible with flow cytometry (FCM), thereby increasing cell detection and enumeration speed by up to two orders of magnitude over microscopic techniques. Moreover, the use of primary Ab conjugated magnetic nanoparticles showed better correlation between epifluorescent microscopy counts and FCM analysis than that achieved using traditional centrifugation techniques. The results strongly support the applicability of the flow-cytometric IMS for microbial detection in complex matrices.

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Acknowledgements

The authors would like to thank Dr. Karlin M. Danielsen and Dr. Kim F. Hayes for providing MNPs. This research is partly funded by a scholarship to Shu-Chi Chang (No. 1999049) from the Ministry of Education, Taiwan, Republic of China.

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  1. Department of Civil and Environmental Engineering, University of Michigan, Room 116, EWRE Bldg., 1351 Beal Avenue, Ann Arbor, MI, 48109, USA

    Shu-Chi Chang, Tracey I. Anderson, Sarah E. Bahrman, Anna I. Khijniak & Peter Adriaens

  2. Department of Civil Engineering, University of Toledo, Toledo, OH, USA

    Cyndee L. Gruden

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  1. Shu-Chi Chang
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  2. Tracey I. Anderson
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Correspondence to Peter Adriaens.

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Chang, SC., Anderson, T.I., Bahrman, S.E. et al. Comparing recovering efficiency of immunomagnetic separation and centrifugation of mycobacteria in metalworking fluids. J IND MICROBIOL BIOTECHNOL 32, 629–638 (2005). https://doi.org/10.1007/s10295-005-0238-x

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