Immunomagnetic nanobeads based on a streptavidin-biotin system for the highly efficient and specific separation of Listeria monocytogenes

Immunomagnetic separation was used to decrease the assay time for Listeria monocytogenes (LM) isolation. The coupling conditions and immunomagnetic bead (IMB) capture process were optimized. Results showed that the incubation time and separation time for 0.1 mg of 180 nm immunomagnetic nanobeads (IMB-S-180) coupled with monoclonal antibody using a streptavidin-biotin system was 45 and 3 min, respectively. Magnetic separation was affected by the separation medium and magnetic density of the magnetic separator. Capture efficiency (CE) was 94.12% when the magnetic density was 1.5 T, using 1 ml of PBS as separation medium. The IMBs exhibited highly specific binding with LM strains (CE > 90%) but low binding with non-target bacteria (CE < 5%). The CE of IMBs (0.1 mg/ml) against LM in ground beef and milk samples were 25.1% and 91.2%, respectively. These results can serve as a guide for the appropriate selection of IMBs for developing isolation processes for pathogens.

[1]  J. Odumeru,et al.  Evaluation of immunomagnetic separation in combination with ALOA Listeria chromogenic agar for the isolation and identification of Listeria monocytogenes in ready-to-eat foods. , 2010, Journal of microbiological methods.

[2]  Il-Hoon Cho,et al.  In situ immuno-magnetic concentration-based biosensor systems for the rapid detection of Listeria monocytogenes , 2012 .

[3]  Weihua Lai,et al.  Development of an immunomagnetic separation method for efficient enrichment of Escherichia coli O157:H7 , 2014 .

[4]  William W. Yu,et al.  Food related applications of magnetic iron oxide nanoparticles: Enzyme immobilization, protein purification, and food analysis , 2012 .

[5]  Zhu Fu,et al.  Rapid detection of Escherichia coli O157:H7 by immunomagnetic separation and real-time PCR. , 2005, International journal of food microbiology.

[6]  Yanbin Li,et al.  Magnetic nanoparticle-antibody conjugates for the separation of Escherichia coli O157:H7 in ground beef. , 2005, Journal of food protection.

[7]  Milica Radisic,et al.  Micro- and nanotechnology in cell separation , 2006, International journal of nanomedicine.

[8]  Lee-Ann Jaykus,et al.  Bacterial Separation and Concentration from Complex Sample Matrices: A Review , 2004, Critical reviews in microbiology.

[9]  D. Kelland.  Magnetic separation of nanoparticles , 1998 .

[10]  Snehal R. Jadhav,et al.  Methods used for the detection and subtyping of Listeria monocytogenes. , 2012, Journal of microbiological methods.

[11]  Haibo Huang,et al.  Magnetic Nanoparticle Based Magnetophoresis for Efficient Separation of E. coli O157:H7 , 2011 .

[12]  Thomas Egli,et al.  Rapid and quantitative detection of Legionella pneumophila applying immunomagnetic separation and flow cytometry , 2010, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[13]  A. Abe,et al.  Combination of immunomagnetic separation with flow cytometry for detection of Listeria monocytogenes. , 2006, Analytica chimica acta.

[14]  Fu-Hsiang Ko,et al.  Rapid and sensitive detection of rare cancer cells by the coupling of immunomagnetic nanoparticle separation with ELISA analysis , 2012, International journal of nanomedicine.

[15]  Weihua Lai,et al.  Establishing of a Method Combined Immunomagnetic Separation with Colloidal Gold Lateral Flow Assay and Its Application in Rapid Detection of Escherichia coli O157:H7 , 2013 .

[16]  Xiuping Jiang,et al.  Rapid detection of Listeria monocytogenes by nanoparticle-based immunomagnetic separation and real-time PCR. , 2007, International journal of food microbiology.

[17]  Mayra Granda Valdés,et al.  Analytical nanotechnology for food analysis , 2009 .

[18]  Jing Xu,et al.  Establishment of magnetic beads-based enzyme immunoassay for detection of chloramphenicol in milk. , 2012, Food chemistry.

[19]  K. Jinneman,et al.  Detection and isolation of low levels of E. coli O157:H7 in cilantro by real-time PCR, immunomagnetic separation, and cultural methods with and without an acid treatment. , 2012, Journal of food science.

[20]  Yanbin Li,et al.  Rapid, sensitive, and simultaneous detection of three foodborne pathogens using magnetic nanobead-based immunoseparation and quantum dot-based multiplex immunoassay. , 2011, Journal of food protection.

[21]  Fei Liu,et al.  Efficient bacterial capture with amino acid modified magnetic nanoparticles. , 2014, Water research.

[22]  Y. Ying,et al.  Evaluation of different micro/nanobeads used as amplifiers in QCM immunosensor for more sensitive detection of E. coli O157:H7. , 2011, Biosensors & bioelectronics.

[23]  A. Wolff,et al.  Isolation and detection of Campylobacter jejuni from chicken fecal samples by immunomagnetic separation–PCR , 2012 .

[24]  P Kolb,et al.  Energy landscape of streptavidin-biotin complexes measured by atomic force microscopy. , 2000, Biochemistry.

[25]  I. Peiris.  Listeria monocytogenes, a Food-Borne Pathogen , 1991, Microbiological reviews.

[26]  J. T. Mayo,et al.  Low-Field Magnetic Separation of Monodisperse Fe3O4 Nanocrystals , 2006, Science.