A novel rapid methodology for determining the chemical oxygen demand (COD) based on a thermal sensor with a flow injection analysis system was proposed and experimentally validated. The ability of this sensor to detect and monitor COD was based on the degree of enthalpy increase when sodium hypochlorite reacted with the organic content in water samples. The measurement results were correlated with COD and were compared against the conventional method using potassium dichromate. The assay required only 5–7 min rather than the 2 h required for evaluation by potassium dichromate. The linear range was 5–1000 mg/L COD, and the limit of detection was very low, 0.74 mg/L COD. Moreover, this method exhibited high tolerance to chloride ions; 0.015 mol/L chloride ions had no influence on the response. Finally, the sensor was used to detect the COD of different water samples; the results were verified by the standard dichromate method.
1 September 2013 in Clinical Microbiology and Infection
There is a need for analytical methods capable of monitoring blood antibiotic levels in real time. Here we present a method for quantifying antibiotic levels in whole blood that does not require any sample pretreatment. The tests employ the enzyme penicillinase to assay for penicillin G, penicillin V and ampicillin using a flow-injected biosensor, the Enzyme Thermistor. Optimal flow rates, sample volumes and pH were determined to be 0.5 mL/min, 100 μL and 7.0, respectively. Analysis of the antibiotics diluted in buffer gave a linear range of 0.17–5.0 mM. Calibration curves prepared using blood spiked with the antibiotics gave a linear range of 0.17–2.0 mM. Linear regression values for all of the calibration curves were 0.998 or higher. Assay cycle time was 5 min. The relative standard deviation value for 100 determinations of a mock blood sample spiked with penicillin G was 6.71%. Despite the elimination of sample pretreatment, no detectable clogging or signal drift was observed. The assay provides a fast, simple, reliable analytical method for determining antibiotic concentrations in blood without the need for any sample pretreatment. This is an important first step towards developing a device capable of real-time monitoring of antibiotic levels in whole blood. The technology has the potential to significantly improve the outcomes of patients undergoing critical care.
Antimicrobial resistance (AMR) threatens global public health and modern surgical medicine. Expression of β-lactamase genes is the major mechanism by which pathogens become antibiotic resistant. Pathogens expressing extended spectrum β-lactamases (ESBL) and carbapenemases (CP) are especially difficult to treat and are associated with increased hospitalization and mortality rates. Despite considerable effort, identification of ESBLs and CPs in a clinically relevant timeframe remains challenging. In this study, a two-dimensional AMR profiling assay strategy was developed employing panels of antibiotics (penicillins, cephamycins, oximino-cephalosporins and carbapenems) and β-lactamases inhibitors (avibactam and EDTA). The assay required the development of a novel biosensor that employed New Delhi metallo-β-lactamase-1 (NDM-1) as the sensing element. Functionally probing β-lactamase activity using substrates and inhibitors combinatorically increased the informational content that enabled the development of assays capable of simultaneous, differential identification of multiple β-lactamases expressed in a single bacterial isolate. More specifically, the assay enabled the simultaneous identification of ESBL and CP in mock samples, as well as in an engineered construct which co-expressed these β-lactamases. The NDM-1 biosensor assay was 16 times and 8 times more sensitive than the ESBL Nordmann/Dortet/Poirel (NDP) and Carba Nordmann/Poirel (NP) assays, respectively. In a retrospective study, NDM-1 biosensor assays were able to differentially identify ESBLs, metallo-CPs and serine-CPs β-lactamases in 23 clinical isolates with 100% accuracy. An assay algorithm was developed which accelerated data analytics reducing turnaround to <1 h. The assay strategy integrated with AI-based data analytics has the potential to provide physicians with a comprehensive readout of patient AMR status.
Real-time monitoring of patient’s blood metabolites, such as glucose and lactate, could potentially improve surgery and recovery outcomes for patients in surgical and intensive care units. Our enzyme thermometric biosensor which is based on flow injected calorimetric determination of immobilized enzyme reaction is capable of performing continuous, fast, and quantitative analysis of metabolites using whole blood. A key technical advantage the assay affords is the ability to use unpretreated whole blood. In this article, the enzyme thermometric biosensor was used, for the first time, to determine glucose and lactate concentrations in the blood of ICU patients. The linear detection range for glucose was 0.5–30 mM and 0.25–12 mM for lactate, using a 20 μL sample volume. A maximum sampling rate of 15 measurements per hour was achieved using venous blood samples, which corresponds to a 4-min measurement interval. In order to validate the accuracy of the results, a comparative analysis between the thermometric biosensor and the clinically applied instrument (LifeScan’s OneTouch®) which is based on disposable dry chemical reaction was performed using samples from 33 patients. The results showed a good correlation between the two methods for both glucose (r = 0.843, p < 0.0001) and lactate (r = 0.78, p = 0.0105). The ability to monitor metabolite levels and trends on a clinically relevant timescale of 5 min is critical for intensive monitoring of ICP and operative patients.