Separation of Listeria from cheese and enrichment media using antibody-coated microbeads and centrifugation

Kaclíková, Eva; Kuchta, T.; Kay, Helen H.; Gray, D.I.

doi:10.1016/s0167-7012(01)00255-x

Cited by 40 publications

(27 citation statements)

References 5 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Other anti-Listeria antibodies recognize only selected serovars (10,12) or secreted proteins (3,32) and are therefore not useful for IMS. Because of the lack of specificity (4,9,29) and poor separation efficiency and sensitivity (5,7,8,9,29), it appears as if antibodies may not be the best-suited tool for Listeria immobilization and separation.…”

Section: Discussionmentioning

confidence: 99%

“…After optimization of several parameters, recovery rates of 86 to 99% could be obtained for cells of different Listeria strains, even in the presence of an excess microbial background. This specificity is mediated by the highly specific recognition properties of the CBDs (15,20) and represents a clear advantage over antibodies, which frequently show cross-reactions with other cells (9,22,29). Important findings were that the target specificity of CBD118 and CBD500 is maintained when the proteins are immobilized on the bead surfaces and that the specific interaction between beads and cells also worked in mixed preparations of CBD118-and CBD500-coated beads.…”

Section: Discussionmentioning

confidence: 99%

“…An ideal tool for the latter appeared to be immunomagnetic separation (IMS). However, these antibody-based anti-Listeria IMS techniques (4,6,9,26,28,29) are hampered by a number of disadvantages, i.e., (i) poor recovery rates-the percentage of cells which could be separated from cell suspensions was 20% or less (9,29), (ii) inability to detect low contamination levels (22,29), and (iii) cross-reaction with nontarget bacterial cells (9,22,29). Moreover, cross-linking and agglutination of beads were frequently observed.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Use of High-Affinity Cell Wall-Binding Domains of Bacteriophage Endolysins for Immobilization and Separation of Bacterial Cells

Kretzer

Lehmann

Schmelcher

et al. 2007

Appl Environ Microbiol

161

136

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Use of High-Affinity Cell Wall-Binding Domains of Bacteriophage Endolysins for Immobilization and Separation of Bacterial Cells

Kretzer

Lehmann

Schmelcher

et al. 2007

Appl Environ Microbiol

161

136

View full text Add to dashboard Cite

“…After the centrifugation, the supernatant is discarded and the bacteria is resuspended in the buffer or broth for further assay or it can be resuspended in buffer with a small volume to achieve the aim of concentration (32). The limitation of the bacterial separation by centrifugation from food containing fat, such as meat or dairy products, is not easy to be achieved.…”

Section: Bacterial Separation From the Liquid Food Matrix 2321 Cenmentioning

confidence: 99%

Enzymatic Digestion for Improved Bacteria Separation from Leafy Green Vegetables

Wang

et al. 2016

Journal of Food Protection

View full text Add to dashboard Cite

An effective and rapid method for the separation of bacteria from food matrix remains a bottleneck for rapid bacteria detection for food safety. Bacteria can strongly attach to a food surface or internalize within the matrix, making their isolation extremely difficult. Traditional methods of separating bacteria from food routinely involve stomaching, blending, and shaking. However, these methods may not be efficient at removing all the bacteria from complex matrices. Here, we investigate the benefits of using enzyme digestion followed by immunomagnetic separation to isolate Salmonella from spinach and lettuce. Enzymatic digestion using pectinase and cellulase was able to break down the structure of the leafy green vegetables, resulting in the detachment and release of Salmonella from the leaves. Immunomagnetic separation of Salmonella from the liquefied sample allowed an additional separation step to achieve a more pure sample without leaf debris that may benefit additional downstream applications. We have investigated the optimal combination of pectinase and cellulase for the digestion of spinach and lettuce to improve sample detection yields. The concentrations of enzymes used to digest the leaves were confirmed to have no significant effect on the viability of the inoculated Salmonella. Results reported that the recovery of the Salmonella from the produce after enzyme digestion of the leaves was significantly higher (P <0.05) than traditional sample preparation methods to separate bacteria (stomaching and manually shaking). The results demonstrate the potential for use of enzyme digestion prior to separation can improve the efficiency of bacteria separation and increase the likelihood of detecting pathogens in the final detection assay.

show abstract

“…Apart from unspecific methods such as filtration or centrifugation, specific methods like immunomagnetic separation using antibody-covered beads have been employed (Besse et al, 2004;Stevens and Jaykus, 2004). Several publications refer to immunomagnetic separation of L. monocytogenes from food after a preenrichment step, but only a few address the direct separation of Listeria cells from food, such as milk or cheese homogenate (Fluit et al, 1993;Nogva et al, 2000;Uyttendaele et al, 2000;Hsih and Tsen, 2001;Kaclíková et al, 2001;Gray and Bhunia, 2005;Yang et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Paramagnetic Beads Coated with RecombinantListeriaPhage Endolysin–Derived Cell-Wall-Binding Domain Proteins for Separation ofListeria monocytogenesfrom Raw Milk in Combination with Culture-Based and Real-Time Polymerase Chain Reaction–Based Quantification

Walcher

Stessl

Wagner

et al. 2010

Foodborne Pathogens and Disease

View full text Add to dashboard Cite

The aim of this study was to evaluate a fast and simple bead-based method using paramagnetic beads covered with recombinant Listeria phage endolysin-derived cell-wall-binding domain proteins specific for Listeria spp. for separation of the foodborne pathogen Listeria monocytogenes from artificially contaminated raw milk. The method was combined with subsequent detection and quantification by the traditional plate-count technique and real-time polymerase chain reaction (PCR). To account for differences in cell properties, recovery rates and the detection limit were determined using five different L. monocytogenes strains for preparation of a 10-fold dilution series in raw milk, spanning an 8-log scale. Two independent test series were performed for each strain, yielding mean recovery rates of 46.6% to 122.8% for detection with the plate-count method, and 64.7% to 95.1% for detection by real-time PCR. A high correlation was found between the number of L. monocytogenes added to the samples and the number of colony forming units recovered by plate count (0.980), as well as the number of bacterial cell equivalents obtained by real-time quantitative PCR (0.987). The detection limit of the combined cell-wall-binding domain proteins/real-time PCR approach ranged from 10(2) to 10(3) colony forming units per milliliter, which is close to the theoretical detection limit of the method.

show abstract

Separation of Listeria from cheese and enrichment media using antibody-coated microbeads and centrifugation

Cited by 40 publications

References 5 publications

Use of High-Affinity Cell Wall-Binding Domains of Bacteriophage Endolysins for Immobilization and Separation of Bacterial Cells

Use of High-Affinity Cell Wall-Binding Domains of Bacteriophage Endolysins for Immobilization and Separation of Bacterial Cells

Enzymatic Digestion for Improved Bacteria Separation from Leafy Green Vegetables

Evaluation of Paramagnetic Beads Coated with RecombinantListeriaPhage Endolysin–Derived Cell-Wall-Binding Domain Proteins for Separation ofListeria monocytogenesfrom Raw Milk in Combination with Culture-Based and Real-Time Polymerase Chain Reaction–Based Quantification

Contact Info

Product

Resources

About