Antibodies and Immunoassays for Detection of Bacterial Pathogens

Banada, Padmapriya P.; Bhunia, Arun K.

doi:10.1007/978-0-387-75113-9_21

Cited by 54 publications

(48 citation statements)

References 182 publications

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“…IMS using PMBs is used to separate and concentrate target pathogens from food samples before detection by plating, immunoassay, PCR, or biosensor methods [31,37,39,42,45,51]. Antibodies [14] or alternative molecules [19,51,52] are used as capture molecules for IMS, and improvements in reagents and assay platform development are essential to enhance assay performance.…”

Section: Discussionmentioning

confidence: 99%

“…L. monocytogenes and L. ivanovii are pathogenic to humans and animals [13]. Many virulence and structural genes or gene products in Listeria could be used as targets for antibody- or nucleic acid-based assay development [14]. L. monocytogenes expresses several virulence proteins [15], including Internalin A (InlA), which promotes bacterial adhesion and invasion of the host cell [15].…”

Section: Introductionmentioning

confidence: 99%

“…Immunological approaches to detect pathogens in food are attractive; however, assay performance depends on the quality and specificity of the antibodies used [14,22]. For detection of Listeria , two types of assay specificities are desired: Listeria genus- or L. monocytogenes -specific tests.…”

Section: Introductionmentioning

confidence: 99%

“…For detection of Listeria , two types of assay specificities are desired: Listeria genus- or L. monocytogenes -specific tests. Anti- Listeria antibodies available from research laboratories or commercial vendors are associated with problems of low affinity [23], reaction to heterologous antigens [24,25], lack of reaction towards all serotypes of L. monocytogenes [23,26-28], lack of reaction due to physiological stress induced by growth media or assay parameters [29,30], and lack of compatibility with certain bioassay platforms [14,22,31]. Thus, there is a need for continued efforts to produce high-quality antibodies.…”

Section: Introductionmentioning

confidence: 99%

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Highly specific fiber optic immunosensor coupled with immunomagnetic separation for detection of low levels of Listeria monocytogenes and L. ivanovii

et al. 2012

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BackgroundImmunomagnetic separation (IMS) and immunoassays are widely used for pathogen detection. However, novel technology platforms with highly selective antibodies are essential to improve detection sensitivity, specificity and performance. In this study, monoclonal antibodies (MAbs) against Internalin A (InlA) and p30 were generated and used on paramagnetic beads of varying diameters for concentration, as well as on fiber-optic sensor for detection.ResultsAnti-InlA MAb-2D12 (IgG2a subclass) was specific for Listeria monocytogenes and L. ivanovii, and p30-specific MAb-3F8 (IgM) was specific for the genus Listeria. At all bacterial concentrations (103–108 CFU/mL) tested in the IMS assay; the 1-μm diameter MyOne beads had significantly higher capture efficiency (P < 0.05) than the 2.8-μm diameter M-280 beads with both antibodies. The highest capture efficiency for MyOne-2D12 (49.2% for 105 CFU/mL) was significantly higher (P < 0.05) than that of MyOne-3F8 (16.6 %) and Dynabeads anti-Listeria antibody (9 %). Furthermore, capture efficiency for MyOne-2D12 was highly specific for L. monocytogenes and L. ivanovii. Subsequently, we captured L. monocytogenes by MyOne-2D12 and MyOne-3F8 from hotdogs inoculated with mono- or co-cultures of L. monocytogenes and L. innocua (10–40 CFU/g), enriched for 18 h and detected by fiber-optic sensor and confirmed by plating, light-scattering, and qPCR assays. The detection limit for L. monocytogenes and L. ivanovii by the fiber-optic immunosensor was 3 × 102 CFU/mL using MAb-2D12 as capture and reporter antibody. Selective media plating, light-scattering, and qPCR assays confirmed the IMS and fiber-optic results.ConclusionsIMS coupled with a fiber-optic sensor using anti-InlA MAb is highly specific for L. monocytogenes and L. ivanovii and enabled detection of these pathogens at low levels from buffer or food.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Highly specific fiber optic immunosensor coupled with immunomagnetic separation for detection of low levels of Listeria monocytogenes and L. ivanovii

et al. 2012

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“…[8][9][10] Many of these antibodies react to antigens that are either shared by pathogenic and nonpathogenic Listeria species or only specific to certain serotypes, or their expression is affected by various environmental factors, such as pH, temperature, and carbon sources. [11][12][13][14][15] Monoclonal antibodies developed to listeriolysin O and phosphatidylcholinespecific phospholipase C from L. monocytogenes showed cross-reaction with L. ivanovii.…”

Section: Lathrop Et Almentioning

confidence: 99%

Pathogen-specific antigen target for production of antibodies produced by comparative genomics

Lathrop¹,

Bailey²,

Kim³

et al. 2014

ANTI

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Listeria monocytogenes continues to be a major public health risk and there is a need for improved rapid detection methods. New and highly specific L. monocytogenes antibodies are needed to advance current detection and meet the needs of industry. This research compared the L. monocytogenes genome with that of L. innocua (a nonpathogenic species of Listeria) and identified nine surface proteins specific to L. monocytogenes. Protein sequences were collected from the database and properties such as hydropathy profile and transmembrane topology were analyzed using TMpred software. Nine peptide sequences were chosen, and synthetic peptides were made and administered to rabbits for antibody production. All nine antibodies were screened against a panel of L. monocytogenes, nonpathogenic Listeria, and nonListeria bacteria. Two of the nine antibodies, ie, the Lm404 and LmC639 polyclonal antibodies, showed a specific reaction to L. monocytogenes internalin B and actin polymerization protein, respectively, and were characterized further by enzyme-linked immunosorbent assay, Western blot, and transmission electron microscopy. In Western blot, both antibodies reacted with the targeted protein and did not cross-react with other Listeria spp. The Lm404 polyclonal antibody showed a high reaction with the panel of 41 L. monocytogenes strains while the LmC639 polyclonal antibody showed a weak reaction. Both the Lm404 and LmC639 polyclonal antibodies showed potential for use in immunoassays for specific detection of L. monocytogenes. This study further indicates that comparative genomics could be used to select pathogen-specific antigen for antibody production.

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Rapid Pathogen Detection Tools

Vongkamjan

Yingkajorn

Thammakulkrajang

2019

Encyclopedia of Analytical Chemistry

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Traditional methods for detection of foodborne pathogenic bacteria in food and clinical samples are typically time‐consuming and require multiple steps or even skilled persons for identification and confirmation of pathogens. Multiple serious pathogens including Escherichia coli, Salmonella spp., Listeria monocytogenes, Campylobacter jejuni, and Vibrio spp. have been linked to foodborne diseases and outbreaks worldwide. Rapid, reliable, and less labor intensive detection methods can simplify the steps for pathogen detection for routine testing or monitoring of food samples as well as for following‐up on foodborne illness cases by testing clinical samples. Monitoring of pathogens can become more common and effortless to perform. Immediate responses to potential pathogen contamination in foods can be one of the most effective ways to control foodborne outbreaks. This section reviews the principles and characteristics of some recent rapid detection methods, including (i) immunoassays and nucleic acid‐based detection platforms and (ii) tools based on metabolites released or consumed. As these methods have gained interest for use to detect pathogens in both food and clinical samples, a perspective on the future directions is also described.

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Antibodies and Immunoassays for Detection of Bacterial Pathogens

Cited by 54 publications

References 182 publications

Highly specific fiber optic immunosensor coupled with immunomagnetic separation for detection of low levels of Listeria monocytogenes and L. ivanovii

Highly specific fiber optic immunosensor coupled with immunomagnetic separation for detection of low levels of Listeria monocytogenes and L. ivanovii

Pathogen-specific antigen target for production of antibodies produced by comparative genomics

Rapid Pathogen Detection Tools

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