An improved amperometric immunosensor for the detection and enumeration of protein A-bearing Staphylococcus aureus

Mirhabibollahi, B.; Brooks, Joy L.; Kroll, R.G.

doi:10.1111/j.1472-765x.1990.tb00138.x

Cited by 28 publications

(9 citation statements)

References 6 publications

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“…However, rapidity and ease of operation are very often traded-off against high sensitivity. Indeed, in most cases described, some pre-enrichment step, usually of 18 hours duration, is essential to increase the number of target organisms to a detectable level (30)(31). The immunelectrodes described in this paper are capable of direct detection of very low numbers of organisms, employing a simple, automatable and rapid procedure.…”

Section: Discussionmentioning

confidence: 98%

Immunoelectrodes for the Detection of Bacteria

Rishpon

Gezundhajt

Soussan

et al. 1992

ACS Symposium Series

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This work describes a rapid and sensitive method for the determination of bacteria. The method is based on an enzyme-taggedimmuno-electrochemical assay. Antibodies are immobilized on disposable carbon felt disc electrodes and are used to capture antigens in test solutions. After a short incubation with a second antibody, which is labeled with the enzyme alkaline phosphatase, the activity of the enzyme electrode thus formed is measured. This enzyme reacts with the substrate p-aminophenyl phosphate and the product of this enzymatic reaction, p-aminophenol, is detected amperometrically. The use of rotating electrodes significantly shortens the incubation times and, together with the computerized electrochemical system, results in extremely high sensitivity. The results obtained with Staphylococcus aereus and Escherichia coli cells show that the system can detect as low as 10 cell/ml in less than 10 minutes.There is a great need for fast and reliable devices capable of detection and identification of various types of bacteria. Traditional microbiological methods are based on cultural techniques which are tedious and slow. A considerable effort has been invested in development of detection methods based on immunological interactions. These methods, which generally involve labeling of antibodies with radioactive, fluorescent or enzymatic label, have been shown to be quite effective, giving specific and quantitative detection of target antigens. During the last decade several attempts were made to combine the specificity of antibody-antigen interaction with the high sensitivity, wide dynamic range, and simplicity of electroanalytical methods. These attempts led to the development of electrochemical immunosensors, several of which detect the corresponding antigen at extremely high sensitivity (1-5).

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Section: Discussionmentioning

confidence: 98%

Immunoelectrodes for the Detection of Bacteria

Rishpon

Gezundhajt

Soussan

et al. 1992

ACS Symposium Series

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“…Up to date, very few amperometric immunosensors have been reported for protein A-bearing S. aureus. Mirhabibollahi et al described immunosensing assays based on a direct electrochemical detection of phenol produced by alkaline phosphatase from phenylphosphate [9,10]. Phenol was detected at a Pt working electrode at a potential of þ 870 mV vs. Ag/AgCl.…”

Section: Introductionmentioning

confidence: 99%

Development of an Amperometric Immunosensor for the Quantification of Staphylococcus aureus Using Self‐Assembled Monolayer‐Modified Electrodes as Immobilization Platforms

et al. 2007

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Amperometric immunosensors for the detection and quantification of S. aureus using MPA self-assembled monolayer modified electrodes for the immobilization of the immunoreagents are reported. Two different immunosensor configurations were compared. A competitive mode, in which protein A-bearing S. aureus cells and antiRbIgG labeled with horseradish peroxidase (HRP) compete for the binding sites of RbIgG immobilized onto the 3-mercaptopropionic acid (MPA) modified electrode, was evaluated. Moreover, a sandwich configuration in which S. aureus cells were immobilized onto the MPA SAM, and RbIgG and antiRbIgG labeled with HRP were further linked to the electrode surface, was also tested. In both cases, TTF was used as the redox mediator of the HRP reaction with H 2 O 2 , and it was co-immobilized onto the MPA-modified gold electrode. After optimization of the working variables for both configurations, the analytical performance of the amperometric measurements carried out at 0.00 V (vs. Ag/ AgCl) showed that the competitive immunosensor exhibited a lower limit of detection (1.6 Â 10 5 S. aureus cells mL À1 ), as well as a better repeatability and reproducibility of the measurements.

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“…It has been previously reported [18][19][20], that the amount of ProtA expressed by S. aureus is proportional to the cell count in the sample. Consequently, calibration curves for S. aureus, expressed as the number of S. aureus attenuated and non-attenuated cells per milliliters, were also constructed.…”

Section: Analytical Characteristics Of the Developed Magnetoimmunosensormentioning

confidence: 96%

“…Following this direction, a few immunosensors have been described for the detection of either ProtA or S. aureus bacteria. Electrochemical transduction systems used have been either amperometric [18][19][20][21][22][23][24][25][26][27], piezoelectric [28] or impedimetric [29]. Still, the sensitivity achieved with most of these methods does not always reach the maximum allowed limit legislated for S. aureus in foodstuffs (absence, 10 or 10 2 cfu g −1 , these values depending on the nature of the foodstuff [30,31]).…”

Section: Introductionmentioning

confidence: 99%

Sensitive and rapid amperometric magnetoimmunosensor for the determination of Staphylococcus aureus

et al. 2012

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The preparation and characteristics of a disposable amperometric magnetoimmunosensor, based on the use of functionalized magnetic beads (MBs) and gold screen-printed electrodes (Au/SPEs), for the specific detection and quantification of Staphylococcal protein A (ProtA) and Staphylococcus aureus (S. aureus) is reported. An antiProtA antibody was immobilized onto ProtA-modified MBs, and a competitive immunoassay involving ProtA antigen labelled with HRP was performed. The resulting modified MBs were captured by a magnetic field on the surface of tetrathiafulvalene-modified Au/SPEs and the amperometric response obtained at -0.15 V vs the silver pseudo-reference electrode of the Au/SPEs after the addition of H2O2 was used as transduction signal. The developed methodology showed very low limits of detection (1 cfu S. aureus/mL of raw milk samples), and a good selectivity against the most commonly involved foodborne pathogens originating from milk. These features, together with a short analysis time (2 h), the simplicity, and easy automation and miniaturization of the required instrumentation make the developed methodology a promising alternative in the development of devices for on-site analysis.

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An improved amperometric immunosensor for the detection and enumeration of protein A-bearing Staphylococcus aureus

Cited by 28 publications

References 6 publications

Immunoelectrodes for the Detection of Bacteria

Immunoelectrodes for the Detection of Bacteria

Development of an Amperometric Immunosensor for the Quantification of Staphylococcus aureus Using Self‐Assembled Monolayer‐Modified Electrodes as Immobilization Platforms

Sensitive and rapid amperometric magnetoimmunosensor for the determination of Staphylococcus aureus

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