Evaluation of Antibodies for Immunomagnetic Separation Combined with Flow Cytometry Detection of Listeria monocytogenes

Four polyclonal anti-Listeria antibodies were evaluated for the detection of Listeria monocytogenes in direct and indirect assays using immunomagnetic separation with flow cytometry. The efficiency of immunocapturing using magnetic beads was also determined. None of the tested antibodies exhibited sufficient specificity or avidity to allow sufficient separation and detection of L. monocytogenes for a useful test that differentiated between negative (without cell) and positive (with cell) samples. Plating results confirmed that cells were captured with Dynabeads anti-Listeria and magnetic beads coated with goat anti-Listeria antibody with recovery ranging from 7 to 23%. Fluorescent-labeled polyclonal antibodies used in this study were not sufficiently specific to allow the detection of L. monocytogenes cells captured by the beads.

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Immunomagnetic Separation and Flow Cytometry for Rapid Detection of Escherichia coli O157:H7

Journal of Food Protection ◽

10.4315/0362-028x-61.7.812 ◽

1998 ◽

Vol 61 (7) ◽

pp. 812-816 ◽

Cited By ~ 40

Author(s):

K. H. SEO ◽

R. E. BRACKETT ◽

J. F. FRANK ◽

S. HILLIARD

Keyword(s):

Escherichia Coli ◽

Flow Cytometry ◽

Magnetic Beads ◽

Fluorescent Antibody ◽

Potassium Phosphate ◽

Immunomagnetic Separation ◽

Escherichia Coli O157 ◽

E Coli ◽

Natural Flora ◽

Short Time

A rapid method for detecting Escherichia coli O157:H7 combining immunomagnetic beads (IMB) and flow cytometry was developed. Labeling antigens separated by IMB with fluorescent antibody enabled the detection of <103 CFU bacteria per ml in pure culture. The optimum concentration of magnetic beads for flow cytometry was lower (ca. 105 particles per ml) than that reported for conventional IMB assay (more than 6 × 106 to 8 × 106 particles per ml). Immunomagnetic separation and flow cytometry (IMFC) were evaluated for detecting E. coli O157:H7 in the presence of a competing microorganism and for detecting antibodies in potassium phosphate buffer. The total assay time from separating antigens with IMB to analyzing with flow cytometry was about 1 h. IMFC detected 103 to 104 CFU of E. coli O157:H7 per ml in ground beef enrichment broth and could effectively discriminate between E. coli O157:H7 and competing natural flora. The new assay system provides another approach to separation and detection of low populations of pathogens and shows potential for detecting low concentrations of toxins and other soluble antigens directly from food in a short time.

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Polyamidoamine (PAMAM) dendrimer-mediated biotin amplified immunomagnetic separation method coupled with flow cytometry for viable Listeria monocytogenes detection

Sensors and Actuators B Chemical ◽

10.1016/j.snb.2017.10.152 ◽

2018 ◽

Vol 257 ◽

pp. 286-294 ◽

Cited By ~ 15

Author(s):

Fulai Li ◽

Fan Li ◽

Zoraida P. Aguilar ◽

Yonghua Xiong ◽

Hengyi Xu

Keyword(s):

Flow Cytometry ◽

Listeria Monocytogenes ◽

Immunomagnetic Separation ◽

Pamam Dendrimer ◽

Separation Method

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Combination of immunomagnetic separation with flow cytometry for detection of Listeria monocytogenes

Analytica Chimica Acta ◽

10.1016/j.aca.2006.03.022 ◽

2006 ◽

Vol 573-574 ◽

pp. 158-163 ◽

Cited By ~ 52

Author(s):

Kyoko Hibi ◽

Akihisa Abe ◽

Eiji Ohashi ◽

Kohji Mitsubayashi ◽

Hideki Ushio ◽

...

Keyword(s):

Flow Cytometry ◽

Listeria Monocytogenes ◽

Immunomagnetic Separation

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Rapid and highly sensitive detection of Flavobacterium psychrophilum using high gradient immunomagnetic separation with flow cytometry

Aquaculture ◽

10.1016/j.aquaculture.2010.09.023 ◽

2010 ◽

Vol 309 (1-4) ◽

pp. 125-130 ◽

Cited By ~ 11

Author(s):

Uni Ryumae ◽

Kyoko Hibi ◽

Yasutoshi Yoshiura ◽

Huifeng Ren ◽

Hideaki Endo

Keyword(s):

Flow Cytometry ◽

Immunomagnetic Separation ◽

Sensitive Detection ◽

High Gradient ◽

Flavobacterium Psychrophilum ◽

Highly Sensitive ◽

Highly Sensitive Detection

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Rapid Flow Cytometry Detection of a Single Viable Escherichia coli O157:H7 Cell in Raw Spinach Using a Simplified Sample Preparation Technique

Frontiers in Microbiology ◽

10.3389/fmicb.2017.01493 ◽

2017 ◽

Vol 8 ◽

Cited By ~ 6

Author(s):

Anna J. Williams ◽

Willie M. Cooper ◽

Shawn Ramsaroop ◽

Pierre Alusta ◽

Dan A. Buzatu ◽

...

Keyword(s):

Escherichia Coli ◽

Flow Cytometry ◽

Sample Preparation ◽

Escherichia Coli O157 ◽

Preparation Technique ◽

Sample Preparation Technique ◽

Rapid Flow ◽

Flow Cytometry Detection

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Can Serum Thymidine Kinase 1 Detect Small Invisible Malignant Tumours?

10.21203/rs.3.rs-902717/v1 ◽

2021 ◽

Author(s):

Ji Zhou ◽

Huijun Li ◽

Cong Fang ◽

Junye Tan ◽

Peng Gao ◽

...

Keyword(s):

Thymidine Kinase ◽

Total Protein ◽

Magnetic Beads ◽

Polyclonal Antibodies ◽

Malignant Tumour ◽

The Body ◽

Cell Diameter ◽

Malignant Tumours ◽

Thymidine Kinase 1 ◽

Protein Cell

Abstract Objectives. Early detection of malignant tumour is a prerequisite for a successful treatment. Here we investigate if thymidine kinase 1 is more sensitive than imaging technology to discover small invisible malignant tumours.Material and Methods. The cellular concentration of TK1 was determined by a novel automatic chemiluminescence analyzer of magnetic particle immune sandwich minimum. The primary and secondary antibodies linked to the magnetic beads were chicken anti-human thymidine kinase 1 IgY-polyclonal antibodies (IgY pAb). The minimum number of cells able to be detected by the novel detection technology using an automatic chemiluminescence analyzer were determined based on the cellar TK1 concentration of low and high TK1 cell lines of known cell count.Results. The TK1 concentration of malignant cell was found to be 0.021 pg/cell. Assuming 200 pg of total protein/cell, TK1 corresponds to 0.01 % of the total protein/cell. The concentration of TK1 in human blood serum of malignant patients is in the range of 2-10 pmol/l (pM), corresponding to about 50 x106 growing cells in the body that release TK1 into 5 litre blood. The limit visibility by imaging of a tumour is about 1 mm in diameter, corresponding to about 109cells of a cell diameter of 1µm. Conclusion. TK1 is more sensitive than imaging.

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