Enhanced Rapidity for Qualitative Detection of Listeria monocytogenes Using an Enzyme-Linked Immunosorbent Assay and Immunochromatography Strip Test Combined with Immunomagnetic Bead Separation

Abstract

An enzyme-linked immunosorbent assay (ELISA), immunochromatography (ICG) strip test, and immunomagnetic bead separation (IMBS) system based on a monoclonal antibody were individually developed for the detection and isolation of Listeria monocytogenes in meat samples. The three methods showed a strong reaction with Listeria species and a weak reaction with Staphylococcus aureus. To increase the rapidity of L. monocytogenes detection, combinations of the ELISA and ICG strip test with the IMBS system (ELISA-IMBS and ICG-IMBS) were investigated. In comparative analyses of artificially inoculated meat and samples of processed meat, the ELISA and ICG strip test required 24 h of enrichment time to detect the inoculated meat samples with > or =1 X 10(2) CFU/10 g, whereas the ELISA-IMBS and ICG-IMBS required only 14 h of enrichment. Analyses of naturally contaminated meat samples (30 pork samples, 20 beef samples, 26 chicken samples, 20 fish samples, and 20 processed meat samples) performed by ELISA-IMBS, ICG-IMBS, and API kit produced similar results. The ELISA-IMBS and ICG-IMBS provide a more rapid assay than the individual ELISA and the ICG strip test and are appropriate for rapid and qualitative detection of L. monocytogenes (or Listeria species) in meat samples. With the ICG-IMBS, L. monocytogenes could be detected in meat samples within 15 h and the method has potential as a rapid, cost-effective on-site screening tool for the detection of L. monocytogenes in food samples and agricultural products at a minimum detection level of approximately 100 CFU/10 g.

Human heat-shock protein 60 receptor-coated paramagnetic beads show improved capture of Listeria monocytogenes in the presence of other Listeria in food

“…2007; Bilir Ormanci et al. 2008), enzyme‐linked immunoassay (Shim et al. 2008), flow cytometry (Jung et al.…”

“…Immunomagnetic separation (IMS) method has been widely used for the capture of varieties of pathogens including L. monocytogenes (Skjerve et al 1990;Benoit and Donahue 2003;Stevens and Jaykus 2004;Bhunia 2008). It has been combined with various assay methods such as isolation on selective or chromogenic agar media (Uyttendaele et al 2000;Kaclikova et al 2001;Bauwens et al 2003;Wadud et al 2010), PCR (Hudson et al 2001;Ueda et al 2006;Yang et al 2007;Bilir Ormanci et al 2008), enzyme-linked immunoassay (Shim et al 2008), flow cytometry (Jung et al 2003;Hibi et al 2006) and cytotoxicity assay (Gray and Bhunia 2005) to precede identification and quantification of L. monocytogenes. The performance of these detection assays can be maximized and improved when combined with a highly efficient magnetic separation method.…”

Development of a monoclonal antibody against deoxynivalenol for magnetic nanoparticle-based extraction and an enzyme-linked immunosorbent assay

“…The binding capacity was 95.44 ± 0.51% and the DON recovery rates for spiked buffer solutions using this novel mAb-MNP system were 75.2, 96.9, and 88.1%. Micronized beads have been widely used as immunosensors for separating glycoproteins, several kinds of cells, bacteria, and DNA from specific matrixes [2,8,21,24,25,27]. However, micro-sized beads have a low dispersion capacity in solution, can make the separation procedure laborious, and produce unreliable data.…”

Rapid and sensitive detection of Salmonella enteritidis by a pre-concentrated immunochromatographic assay in a large-volume sample system

“…17,18 Immunomagnetic separation has been a useful technology to concentrate and separate target pathogens from complex food samples. [19][20][21][22] In recent years, immunomagnetic separation was also coupled with ICA to improve the detection sensitivity and specicity. For example, Cui et al 23 established a method combined immunomagnetic separation with CG-ICA to isolate and detect Escherichia coli O157:H7.…”