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View of amperometric immunosensor for penicillin G determination using an H2O2 electrode (size: ø = 1 cm; width = 7 cm) as transducer and peroxidase as enzyme label.
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The aim of the present research was to develop a single use, simple but highly sensitive amperometric immunosensor for penicillin G and other β-lactam antibiotics based on a “competitive assay”. The immunosensor developed uses an amperometric electrode for hydrogen peroxide as transducer and the peroxidase enzyme as marker. The results demonstrate...
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... immunosensor (Fig. 1) was assembled using an Immobilon membrane in which the antibody or the antigen was immobi- lized and which overlapped a cellulose acetate membrane (0.1-mm thick) placed on the lower end of the plastic cap of an amperometric electrode for H 2 O 2 . A nylon net and a rubber O-ring were used to fix the Immobilon membrane to the head of ...
Citations
... The adequate performance of the sensor can be achieved by optimizing detection parameters. Electrochemical immunosensor designed for detection of penicillin G and other beta-lactam antibiotics river wastewater has also shown high potential for sensitive detection of other antibiotic contaminants including dicloxacillin, amoxicillin, ampicillin, and cefotaxime (Merola et al., 2014). Yang et al. (2017) have explored synthetic aptamers as a receptor for the development of high stability and cost-effective electrochemical aptasensors as an alternative to antibodies-based biosensors. ...
The influx of emerging, prescribed, and FDA approved pharmaceutical contaminants (PCs) in the environment is continuously affecting human health and human cycle (even at trace amount) via contaminating water and aquatic life. Among them, non-degradable water-soluble residues which have been integrated with water streams and groundwater is a serious concern and is now a focus of United Nation's Sustainable Development Goals 2030. In this direction, the removal and eradication of PCs from wastewater is challenging but seems manageable via selective detection and efficient remediation using nano-enabled functional systems. To explore this field well for covering gaps, this article is an attempt to explain wide variety of emerging pharmaceutical contaminants, related bio-toxicity, health risks, detection techniques, and approaches of remediation. With the aim of health and environmental management, selective detection and eradication of PCs simultaneously is recommended using nano-system of tuneable properties managed via adopting green approaches. Significant efforts are being made to achieve 1) detection of PCs at very low level (ppb) needed for early-stage water diagnostics and 2) eradicating PCs using stimuli-responsive catalysis. In both the cases, the design and development of electro-active nano-systems of desired feature is key requirement for developing acceptable and affordable health and environmental management strategies. The outcomes of this article will encourage scholars to develop novel and efficient nano-systems capable of detection and eradication of targeted PCs to produce clean and healthy water for everyone.
... Electrochemical sensors can be a reliable method for on-site detection of pharmaceutical particles, and it can be easily miniaturized and automated. Merola et al. [107] established an electrochemical immune-sensor utilizing two competitive assays for a sensitive detection of penicilin G and other -lactam antibiotics. There are two perspectives allowed by the electrochemical techniques: electrochemical conversion in which the recalcitrant organic pollutants are particularly modified into biodegradable compounds and electrochemical combustion within which organic chemicals are mineralized. ...
A wide range of unregulated chemicals of synthetic origin or derived from natural sources, which may be a contender for future regulations are called Emerging Contaminants (ECs). The concentration of ECs ranges from ng/L to μg/L, which is comparatively smaller as compared to other pollutants present in water and wastewater. Even though the environmental concentration is low, ECs still possess a great threat to the humans and ecosystem. These compounds are being widely studied due to their potential health effects, pervasive nature, and difficult degradation through conventional techniques. Pharmaceutical active compounds (PhACs) or pharmaceutical contaminants (PCs) are one of the major groups of ECs which can cause inimical effect on living organisms even at very lower concentration. These contaminants don't degrade easily and persistent for longer periods in the environment due to their stable structure. With the increase in demand of Pharmaceuticals and Personal Care Products (PPCPs), there has been a sharp increase of these pollutants in water bodies. This is mainly due to the inefficiency of conventional wastewater treatment plants in treatment and removal of these PhACs. The proper identification of pharmaceutical groups and development of removal techniques is crucial in the recent times. This review represents a comprehensive summary on PCs, various groups of PCs and an overview of approaches and treatment systems available for their removal. Efficient and effective treatment methods can be useful for completely eradicating these compounds and making the water bodies safe for use. So, the investment of capital and time on research on PCs and their removal techniques can be beneficial for the future.
... Beta-lactam antibiotics are one of the most widely used classes of antimicrobial drugs in medicine and veterinary chemotherapy [1,2]. Therefore, accurate, simple and rapid methods for their determination are in demand and are relevant for monitoring the biosafety of food of animal origin [3][4][5][6], assessing the level of beta-lactams in the blood in order to prescribe an adequate dose to patients in emergency conditions [6][7][8][9], as well as for monitoring content in environmental objects [10,11]. An immunoassay based on the antigen-antibody reaction has been recognized as an effective method of screening control of various antibiotics [3,[12][13][14][15]. ...
Abstract—Fifteen protein conjugates of penicillins and cephalosporins containing amino- and/or carboxylic groups in the initial structures have been synthesized in the reactions with human serum albumin or ovalbu-min using 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC) or a combination of EDC and N-hydroxysulfosuccinimide at various ratios of the base reagents. A comparative study of conjugates composition and properties has been carried out by UV spectroscopy, mass spectrometry and a ligand-recep-tor assay. It was shown that the antibiotic residue content of the macromolecules obtained varied from 1 to 22, the beta-lactam cycle remained intact assuring specific interactions of the conjugates with a penicillin-binding protein. In two developed models of receptorbioanalytic systems, an ampicillin conjugate onto a
solid phase binds to penicillin-binding protein complexed with a monoclonal antibody, which was detected by an immunoenzyme label in microplate wells or goldnanoparticles on test strips. Conjugated ampicillin binding to the receptor was competitively inhibited by beta-lactam antibiotics added to the liquid phase, and analytical sensitivities relative to penicillin G were 0.05 and 1 ng/mL for microplate and receptor chromato-graphic systems, respectively.
... An over-use of these substances results in their residues to be accumulated in food, surface water, soil, etc., predominantly in animal-derived foods and other dairy products [55,56]. It is extremely important to detect the traces of these contaminants in all kinds of foodstuffs as otherwise they may pose severe health concerns for human beings, ranging from resistant bacterial evolution to allergic reactions in individuals [57] and bacterial resistance related infections [58]. The most important details of the AuNPs based colorimetric aptasensors being discussed for the antibiotics of each group individually, including the aptamer sequences, dissociation constant (KD), the limit of detection (LOD), color change, AuNP particle size, and synthesis protocol are listed in their respective tables for each antibiotic class ( Table 2). ...
Misuse of antibiotics has recently been considered a global issue because of its harmful effects on human health. Since conventional methods have numerous limitations, it is necessary to develop fast, simple, sensitive, and reproducible methods for the detection of antibiotics. Among numerous recently developed methods, aptasensors are fascinating because of their good specificity, sensitivity and selectivity. These kinds of biosensors combining aptamer with colorimetric applications of gold nanoparticles to recognize small molecules are becoming more popular owing to their advantageous features, for example, low cost, ease of use, on-site analysis ability using naked eye and no prerequisite for modern equipment. In this review, we have highlighted the recent advances and working principle of gold nanoparticles based colorimetric aptasensors as promising methods for antibiotics detection in different food and environmental samples (2011–2020). Furthermore, possible advantages and disadvantages have also been summarized for these methods. Finally, the recent challenges, outlook, and promising future perspectives for developing novel aptasensors are also considered.
... The LOD was in the order of 1.0 Â 10 À10 mol L -1 . The immunosensor developed demonstrated low selectivity toward all b -lactam antibiotics, higher selectivity toward other classes of non-blactam antibiotics and good recovery in artificially prepared samples [144]. ...
... A detailed comparison between the different polymers modified electrodes and biosensors in terms of analytical figures of merit is reported in Table 3 [52][53]89,94,96,[133][134][135][136][139][140][141][143][144]146,148]. ...
Amoxicillin (AMX) is among the most successful antibiotics used for human therapy. It is used extensively to prevent or treat bacterial infections in humans and animals. However, the widespread distribution and excess utilization of AMX can be an environmental and health risk due to the hazardous potential associated to its pharmaceutical industries effluents. Besides, their extensive use in food animal production may result in some undesirable residues in food, e.g. meat, eggs and milk. Consequently, at high enough concentrations in biological fluids, AMX may be responsible of various diseases such as nausea, vomiting, rashes, and antibiotic-associated colitis. For this reason, the detection and quantification of amoxicillin in pharmaceuticals, biological fluids, environmental samples and foodstuffs require new electroanalytical techniques with sensitive and rapid measurement abilities. This review discusses recent advances in the development of electrochemical sensors and bio-sensors for AMX analysis in complex matrices such as pharmaceuticals, biological fluids, environmental water and foodstuffs. The main electrochemical sensors used are based on chemically modified electrodes involving carbon materials and nanomaterials, nanoparticles, polymers and biological recognition molecules.
... The detection of traces of the contaminants in food is extremely important, as they can have serious health consequences on humans, which range from allergic reactions in sensitive individuals to the evolution of antibiotic-resistant bacteria [2,5,9,10] and enhanced bacterial resistance infections [11]. ...
This work aimed to develop accurate, quick, and practical tools for the detection of residues of penicillin G antibiotic in biological and non-biological samples. The assays were developed based on the binding mechanism of β-lactam to penicillin-binding proteins; samples of different concentrations of penicillin G were incubated with in vitro expressed 6X-Histidine-tagged soluble penicillin-binding protein (PBP2x*) of Streptococcus pneumoniae (S. pneumoniae), whereby penicillin G in samples specifically binds to PBP2x*. The fluorescent-labeled β-lactam analogue Bocillin FL was used as a competent substrate, and two different routes estimated the amounts of the penicillin G. The first route was established based on the differences in the concentration of non-bounded Bocillin FL molecules within the reactions while using a real-time polymerase chain reaction (PCR)-based method for fluorescence detection. The second route depended on the amount of the relative intensity of Bocillin FL bounded to Soluble PBP-2x*, being run on sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-page), visualized by a ChemiDoc-It®2 Imager, and quantified based on the fluorescence affinity of the competent substrate. While both of the methods gave a broad range of linearity and high sensitivity, the on column based real-time method is fast, non-time consuming, and highly sensitive. The method identified traces of antibiotic in the range 0.01–0.2 nM in addition to higher accuracy in comparison to the SDS-based detection method, while the sensitivity of the SDS-based method ranged between 0.015 and 2 µM). Thus, the on column based real time assay is a fast novel method, which was developed for the first time based on the binding inhibition of a fluorescence competitor material and it can be adapted to screen traces of penicillin G in any biological and environmental samples.
... Technologies in research for detection of antimicrobials include colorimetric [21], surface plasmon resonance (SPR) [22,23], fluorescence spectroscopy [24,25], surface enhanced Raman spectroscopy (SERS) [26], voltammetry [27,28], and piezoelectric based techniques [29]. Most of these technologies are integrated with either natural receptors such as antibodies [30,31] and enzymes [32,33], or synthetic receptors such as molecular imprinted polymers (MIPs) [28,34] and aptamers [35] for antibiotic detection. While sensors utilizing antibodies and enzymes offer high specificity, scalability of such devices remains an issue, mainly due to the costs involved. ...
Antimicrobial resistance affecting humans is fueled by empirical infection diagnostics and continuous unintended antibiotics exposure. A localized surface plasmon resonance (LSPR) based optical fiber biosensor for capture of bacteria followed by antibiotic mediated lysis has been demonstrated for quick drug susceptibility testing. The concept has been proven with P. aeruginosa and E. coli suspended in simulated human urine for potential drug susceptibility testing for urinary tract infections. P. aeruginosa RS1 was found to be selectively sensitive to commonly used third generation cephalosporins ceftazidime, ceftriaxone and cefotaxime. Hence, in an alternate scheme, bacteriolysis signatures of P. aeruginosa immobilized on gold nanoparticles modified optical fibers was used for quantification of third generation cephalosporins over a linear range of 0.01 to 1 μg/ml, R² = 0.985, in water. Further, this principle was established for therapeutic drug monitoring (TDM) of the said antibiotics in real human serum in the therapeutic relevant range of 0.5 to 10 µg /mL (R² = 0.987). This sensor provides a quick alternative to cumbersome Kirby-Bauer disk diffusion tests, practiced in most community health settings and tertiary care hospitals for drug susceptibility testing. The sensor also provides an alternative to chromatography coupled mass spectroscopic techniques for quicker, affordable and possibly point of care antibiotic quantification with applications in environmental and therapeutic monitoring.
... Among the various kinds, great interest is attached to competitive immunosensor due to its high sensitivity for detection of small molecules. Competitive immunosensors have been developed to analyze small analyses such as 2,4-dinitrophenol (Aizawa et al. 2007), pesticides (Schipper et al. 1997), antibiotics (Merola et al. 2014), and aflatoxin B1 (Majer-Baranyi et al. 2016) at low detection limit. ...
Tyramine is a naturally occurring monoamine compound produced from decarboxylation of the amino acid tyrosine by monoamine oxidase. Foodstuffs containing considerable amounts of tyramine include fish, fermented food, and other dairy products. High levels of tyramine in food samples cause toxic effects such as migraine, tachycardia, and vomiting. Therefore, sensitive methods are required to monitor and detect tyramine. Here, a competitive colorimetric immunosensor was developed for ultrasensitive detection of tyramine in fish samples. Tyramine-bovine serum albumin was coated on a microplate and introduced as an analyze competitor. Competitive assays were achieved by incubating free tyramine and horseradish peroxidase (HRP)–labeled monoclonal anti-tyramine antibodies. After the competitive reaction, captured HRP catalyzed the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) in the presence of hydrogen peroxide (H2O2) to produce color responses. The competitive colorimetric immunosensor showed good performance with low detection limit (2 ng/mL), wide linear range (from 5 to 40 ng/mL), high precision (% RSD = 3), and good selectivity in tyramine detection. Applicability of the proposed immunosensor was evaluated by analyzing tyramine in fish samples. Detection of tyramine in real samples showed good recovery and corroboration with the HPLC method. Results indicated that this method can be successfully used for analysis and safety control of tyramine in food samples.
... Merola with his coworkers reported another immunodevice for penicillin G determination [87]. The authors tested two different competitive immunoassays employing an antibiotic or an antibody conjugation with a HRP enzyme. ...
Antibiotics are an important class of drugs destined for treatment of bacterial diseases. Misuses and overuses of antibiotics observed over the last decade have led to global problems of bacterial resistance against antibiotics (ABR). One of the crucial actions taken towards limiting the spread of antibiotics and controlling this dangerous phenomenon is the sensitive and accurate determination of antibiotics residues in body fluids, food products, and animals, as well as monitoring their presence in the environment. Immunosensors, a group of biosensors, can be considered an attractive tool because of their simplicity, rapid action, low-cost analysis, and especially, the unique selectivity arising from harnessing the antigen–antibody interaction that is the basis of immunosensor functioning. Herein, we present the recent achievements in the field of electrochemical immunosensors designed to determination of antibiotics.
... Ampicillin, like kanamycin, is a commonly-used antibiotic. However, ampicillin belongs to the -lactam group, with a mechanism of action similar to penicillin [33][34][35] . Unlike most other penicillins, it is able to be adequately absorbed with oral doses while maintaining low-toxicity in humans and rats 33 . ...
... As an emerging contaminant, the detection antibiotics in wastewater and river water has been a common subject of research. Two common types of antibiotics searched for are thelactams and the aminoglycoside families, due to their widespread use in prescription medicine and in agriculture 34,[64][65][66] . Methods for detection vary, but most involve low-temperature chromatography and different types of spectroscopic measurement 26,27,[65][66][67][68][69] . ...
... Methods for detection vary, but most involve low-temperature chromatography and different types of spectroscopic measurement 26,27,[65][66][67][68][69] . Enzyme assays and indicator tests are also available with varying degrees of accuracy 28,34,64,70,71 . While these tests are able to detect concentrations of the antibiotics to low concentrations (9 ng/L or 9E-10 wt % in water, 2 pg with SERS, and in the µM range over all other media), most of the tests are not portable, and the ones that are either require tedious preparation or can become costly. ...
Detection of Environmental Contaminants in Water Utilizing Raman Scanning for E. coli Phenotype Changes. Hunter Flick Academic Abstract Raman spectroscopy and its counterpart surface-enhanced Raman scattering (SERS) have proven to be effective methods for detecting miniscule changes in the phenotypes of E. coli and other single-celled organisms to aid in the detection of new strains for industrial use and discovery of new antibiotics. The purpose of this study is to develop a method to quickly and accurately detect contaminants in water samples through phenotype changes in E. coli measured through SERS. Contaminated Luria-Bertani (LB) media was inoculated with LB with an OD600 of 1, grown for two hours, and then dried on a flat piece of aluminum foil. These samples were then Raman scanned and processed to determine contaminant-induced changes to the phenotypes of the E. coli. Three types of tests were run to show the effectiveness of this method: single-component, multicomponent, and impure water sources. In single-component tests, it was found that differences due to NaCl contamination could be detected to 5.0E-9 weight percent (wt %), ethanol (EtOH) to 5.0E-7 volumetric percent (% v/v), citric acid (CA) to 2.8E-4 wt %, acetic acid (AA) to 2.6E-4 wt %, kanamycin to 2.5E-11 wt %, ampicillin to 2.5E-10 wt %, CoCl2 to trace amounts, and silver nanoparticles (AgNP) to 5.2E-7 wt %. Many of these are below the detection limits of analytical instrumentation, but their effects on E. coli phenotypes were detectable by Raman spectroscopy. Multicomponent tests showed that in a mixture, the most toxic or most concentrated contaminants have the most effect on cell phenotype. However, it was shown that similar concentrations of similar contaminants may be difficult to discern with current methods. This behavior was also seen in the impure water samples, showing that tap water behaves the closest to a DI control, followed by running water, and finally stagnant bodies. This new method of monitoring E. coli phenotypes with Raman spectroscopy as a biosensor shows promise for the fast, portable, and accurate determination of environmental contaminants with a broad-spectrum and very low detection limits. Detection of Environmental Contaminants in Water Utilizing Raman Scanning for E. coli Phenotype Changes. Hunter Flick General Audience Abstract Recently, Raman spectroscopy and an enhanced version called surface-enhanced Raman scattering (SERS) have shown promise in showing the effects of a cell's environment on how it expresses genes and what chemical compounds it produces to survive. It does this through reading the chemical signature it gives off due to these changes, and these readings have been used for showing the effects of antibiotics, finding varieties that are resistant to toxic byproducts of their activities, and as biosensors. The study outlined in this thesis aims to develop a test utilizing SERS to see the reactions of a non-pathogenic strain of E. coli to contaminants in their media and determine their identity. This test was run for three types of contaminated samples: one contaminant, three contaminants, and tests using impure water from a sink tap, a pond, and a stream. For the single contaminants, eight types were tested; NaCl, ethanol, citric acid, acetic acid (AA), kanamycin, ampicillin, CoCl2, and silver nanoparticles. Detection limits for all contaminants were found, with the lowest detectable concentrations all falling below or matching detection limits of common methods. The lowest detectable concentrations came from kanamycin and CoCl2, at 2.5E-11 weight percent and in amounts which are considered beneficial to the environment, respectively. The three-contaminant test shows that it is possible to pinpoint which contaminants are having the highest effect, though if contaminants are similar in nature and in similar concentrations, it may be difficult to pinpoint which is causing the effect. In the final test, similarity of water sources to pure water were found, with tap water being closest, followed by stream water and the most different being pond water. It was also found that pond water and stream water were closest in behavior, showing the power of this method in differentiating sources from each other. iv
