Electrochemical sensors for identifying pyocyanin production in clinical Pseudomonas aeruginosa isolates

Sismaet, Hunter J.; Pinto, Ameet J.; Goluch, Edgar D.

doi:10.1016/j.bios.2017.05.042

Cited by 63 publications

(65 citation statements)

References 32 publications

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“…The wavelength of 600 nm was chosen for OD, because 600 nm does not interfere with the absorbance of GFP, which absorbs at 475 nm. Therefore, relative errors are eliminated by avoiding interfering wavelengths between bacteria and fluorescent proteins or other fluorescent molecules that are present in PA, including pyocyanine and pyoverdine [53,54]. This point also strengthens the argument regarding why additional methods for bacterial quantification, such as fluorescence spectroscopy using EGFP, are desired, as they avoid the overlap of absorbing wavelengths.…”

Section: Fluorescence Spectroscopy As a Tool To Quantify The Planktonmentioning

confidence: 58%

Using Fluorescence Intensity of Enhanced Green Fluorescent Protein to Quantify Pseudomonas aeruginosa

et al. 2018

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A variety of direct and indirect methods have been used to quantify planktonic and biofilm bacterial cells. Direct counting methods to determine the total number of cells include plate counts, microscopic cell counts, Coulter cell counting, flow cytometry, and fluorescence microscopy. However, indirect methods are often used to supplement direct cell counting, as they are often more convenient, less time-consuming, and require less material, while providing a number that can be related to the direct cell count. Herein, an indirect method is presented that uses fluorescence emission intensity as a proxy marker for studying bacterial accumulation. A clinical strain of Pseudomonas aeruginosa was genetically modified to express a green fluorescent protein (PA14/EGFP). The fluorescence intensity of EGFP in live cells was used as an indirect measure of live cell density, and was compared with the traditional cell counting methods of optical density (OD 600 ) and plate counting (colony-forming units (CFUs)). While both OD 600 and CFUs are well-established methods, the use of fluorescence spectroscopy to quantify bacteria is less common. This study demonstrates that EGFP intensity is a convenient reporter for bacterial quantification. In addition, we demonstrate the potential for fluorescence spectroscopy to be used to measure the quantity of PA14/EGFP biofilms, which have important human health implications due to their antimicrobial resistance. Therefore, fluorescence spectroscopy could serve as an alternative or complementary quick assay to quantify bacteria in planktonic cultures and biofilms.

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Section: Fluorescence Spectroscopy As a Tool To Quantify The Planktonmentioning

confidence: 58%

Using Fluorescence Intensity of Enhanced Green Fluorescent Protein to Quantify Pseudomonas aeruginosa

et al. 2018

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“…An actual electrochemical detection of pyocyanin in clinical isolates was conducted by Sismaet et al on 94 different isolates from patients with hospital-acquired infections or patients with cystic fibrosis. The results proved that all the isolates produced pyocyanin [ 42 ]. This is an important finding for the direct identification of PA in clinical samples, as nonproducing PA isolates would lead to false negatives when using pyocyanin as a biomarker for PA.…”

Section: Pyocyanin Detection In Clinical Isolatesmentioning

confidence: 97%

“…To successfully apply the electrochemical detection of pyocyanin for PA identification in clinical samples, several studies measured human fluids spiked with pyocyanin [ 31 , 32 , 48 ]. The investigated human fluids covered urine, blood, bronchial lavage, sputum, saliva and hypertonic saline for laryngeal aspirate suctions as well as simulated wound fluid and artificial sputum medium [ 25 , 33 , 37 , 42 , 43 ]. PA is a urinary tract pathogen, which makes urine an obvious body fluid to develop pyocyanin detection in.…”

Section: Towards Clinical Applicationsmentioning

confidence: 99%

Electrochemical Detection of Pyocyanin as a Biomarker for Pseudomonas aeruginosa: A Focused Review

Alatraktchi

Svendsen

Molin

2020

Sensors

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Pseudomonas aeruginosa (PA) is a pathogen that is recognized for its advanced antibiotic resistance and its association with serious diseases such as ventilator-associated pneumonia and cystic fibrosis. The ability to rapidly detect the presence of pathogenic bacteria in patient samples is crucial for the immediate eradication of the infection. Pyocyanin is one of PA’s virulence factors used to establish infections. Pyocyanin promotes virulence by interfering in numerous cellular functions in host cells due to its redox-activity. Fortunately, the redox-active nature of pyocyanin makes it ideal for detection with simple electrochemical techniques without sample pretreatment or sensor functionalization. The previous decade has seen an increased interest in the electrochemical detection of pyocyanin either as an indicator of the presence of PA in samples or as a tool for quantifying PA virulence. This review provides the first overview of the advances in electrochemical detection of pyocyanin and offers an input regarding the future directions in the field.

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“…Indeed, this virulence factor is uniquely synthesized by PA as it is the only bacterial species carrying the two specific protein‐encoding genes (PhzM, PhzS) necessary to form PYO. [ 62 ] PYO has been proven to exist in all nosocomial strains of PA within a clinically viable concentration of 2–100 µ m , though concentrations of up to 130 µ m have been reported in instances of pulmonary infection, namely bronchiectasis. [ 20b,63 ] Consequently, PYO is of great interest for the detection of PA. PYO presents an oxidized and reduced forms following the gain or loss of electrons at the standard redox potential of −247 mV versus Ag/AgCl (Figure 4C).…”

Section: Electrochemical Detection Of Pamentioning

confidence: 99%

Emerging Technologies for the Electrochemical Detection of Bacteria

McEachern

Harvey

Merle

2020

Biotechnology Journal

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Infections are a huge economic liability to the health care system, although real‐time detection can allow early treatment protocols to avoid some of this cost and patient morbidity and mortality. Pseudomonas aeruginosa (PA) is a drug‐resistant gram‐negative bacterium found ubiquitously in clinical settings, accounting for up to 27% of hospital acquired infections. PA secretes a vast array of molecules, ranging from secondary metabolites to quorum sensing molecules, of which many can be exploited to monitor bacterial presence. In addition to electrochemical immunoassays to sense bacteria via antigen–antibody interactions, PA pertains a distinct redox‐active virulence factor called pyocyanin (PYO), allowing a direct electrochemical detection of the bacteria. There has been a surge of publications relating to the electrochemical tracing of PA via a myriad of novel biosensing techniques, materials, and methodologies. In addition to indirect methods, research approaches where PYO has been sensitively detected using surface modified electrodes are reviewed and compared with conventional PA‐sensing methodologies. This review aims at presenting indirect and direct electrochemical methods currently developed using various surface modified electrodes, materials, and electrochemical configurations on their electrocatalytic effects on sensing of PA and in particular PYO.

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Electrochemical sensors for identifying pyocyanin production in clinical Pseudomonas aeruginosa isolates

Cited by 63 publications

References 32 publications

Using Fluorescence Intensity of Enhanced Green Fluorescent Protein to Quantify Pseudomonas aeruginosa

Using Fluorescence Intensity of Enhanced Green Fluorescent Protein to Quantify Pseudomonas aeruginosa

Electrochemical Detection of Pyocyanin as a Biomarker for Pseudomonas aeruginosa: A Focused Review

Emerging Technologies for the Electrochemical Detection of Bacteria

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