An insight into the state of nanotechnology-based electrochemical biosensors for PCOS detection.

Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorders affecting many women of reproductive age all over the world. PCOS is associated with the onset of enduring health complications, notably diabetes and cardiovascular diseases. Furthermore, PCOS escalates the propensity for conditions such as obesity, insulin resistance, and dyslipidemia, which can potentially culminate in life-threatening scenarios. A pervasive predicament surrounding PCOS pertains to its underdiagnosis due to discrepancies in diagnostic criteria and the intricacy of available testing methodologies. Consequently, many women encounter substantial delays in diagnosis with traditional diagnostic approaches. Prompt identification is imperative, as any delay can precipitate severe consequences. The conventional techniques employed for PCOS detection typically suffer from suboptimal accuracy, protracted assay times, and inherent limitations, thereby constraining their widespread applicability and accessibility. In response to these challenges, various electrochemical methods leveraging nanotechnology have been documented. In this concise review, we endeavor to delineate the deficiencies associated with established conventional methodologies while accentuating the distinctive attributes and benefits inherent to contemporary biosensors. We place particular emphasis on elucidating the pivotal advancements and recent breakthroughs in the realm of nanotechnology-facilitated biosensors for the detection of PCOS.

Advances in surface-enhanced Raman spectroscopy-based sensors for detection of various biomarkers.

Surface enhanced Raman spectroscopy (SERS) allows the ultrasensitive detection of analytes present in traces or even single molecule levels by the generation of electromagnetic fields. It is a powerful vibrational spectroscopic method that is capable to detect traces of chemical and biological analytes. SERS technique is involved in the extremely sophisticated studies of molecules with high specificity and sensitivity. In the vicinity of nanomaterials decorated surfaces, SERS can monitor extremely low concentrations of analytes in a non-destructive manner with narrow line widths. This review article is focused on some recently developed SERS-based sensors for distinct types of analytes like disease-related biomarkers, organic and inorganic molecules, various toxins, dyes, pesticides, bacteria as well as single molecules. This study aims to enlighten the arising sensing approaches based on the SERS technique. Apart from this, some basics of the SERS technique like their mechanism, detection strategy, and involvement of some specific nanomaterials are also highlighted herein. Finally, the study concluded with some discussion of applications of SERS in various fields like food and environmental analysis.

Empowering medicine residents to order labs mindfully to improve patient-centered care.

BACKGROUND: Unnecessary laboratory testing of hospitalized patients is prevalent. OBJECTIVE: We conducted a study focused on "mindful ordering" to decrease unnecessary laboratory ordering within an Internal Medicine residency program. DESIGNS, SETTINGS AND PARTICIPANTS: We collected survey data on resident/faculty perceptions of laboratory ordering as well as order information from the electronic medical record (EMR). INTERVENTION: Interventions focused on resident-identified barriers such as knowledge, EMR, habit and faculty expectations. Interventions were cumulative and included resident/faculty education and EMR optimization. MAIN OUTCOMES AND MEASURES: We assessed basic and complete metabolic panels (BMP, CMP) and complete blood counts with and without differential (CBC w/diff, w/o diff). Primary outcomes included: total labs ordered per week, lab and frequency, and resident perception of ordering practices. Secondary outcomes included: length-of stay (LOS) and venipuncture utilization. RESULTS: Survey data demonstrated increased resident perception of both mindful ordering and team discussion. Total labs ordered per week decreased 20% in the first year (1944 to 1500 labs/week). Residents' use of the "one-time draw" option increased; use of "daily" frequency decreased. Trends showed an increase in BMP relative to CMP, and an increase in CBC w/o diff relative to CBC w/diff. These changes were sustained through 127 weeks. There was an approximately 10% decrease in monthly average of patients undergoing venipuncture each day (86.7% to 74.2%). The shifts in laboratory ordering in conjunction with increased discussion about labs suggest a sustained change in resident lab ordering behavior. This study shows the impact of focusing interventions on resident-identified barriers to mindful ordering to create a sustained decrease laboratory orders.

Electrochemical Sensor for Bilirubin Detection Using Paper-Based Screen-Printed Electrodes Functionalized with Silver Nanoparticles.

A notable diagnostic for the detection of hemolytic diseases is bilirubin, a by-product of haemoglobin breakdown. The concentration of bilirubin ranges from 0.3 to 1.9 mg in 100 mL of blood. Low blood bilirubin levels are associated with a greater risk of coronary heart disease and anaemia. Hyperbilirubinemia results from a serum bilirubin level of more than 2.5 mg/100 mL. Therefore, it is very crucial to check the serum bilirubin level. Analytical equipment for point-of-care testing must be portable, small, and affordable. A unique method is used to detect bilirubin selectively using paper-based screen-printed carbon electrodes that were covalently linked with nanoparticles, that serves as a key biomarker for jaundice. In order to create an electrochemical biosensor, bilirubin oxidase was immobilised on electrodes modified with AgNPs. The morphology of Ag nanoparticles in terms of size and shape was determined using both UV- Vis Spectroscopy and transmission electron microscopy (TEM). The biosensor's analytical response was assessed using potentiostat (Cyclic voltammetry (CV) and linear sweep voltammetry (LSV)). The developed paper-based sensor provided optimum feedback and a broad linear range of 1 to 9 µg/mL for bilirubin, with a lower LOD of 1 µg/mL. Through tests of bilirubin in artificial blood serum, the viability is confirmed. The method that is being used makes it possible to create and use an inexpensive, miniature electrochemical sensor.

Synthesis, characterization and immobilization of bilirubin oxidase nanoparticles (BOxNPs) with enhanced activity: Application for serum bilirubin determination in jaundice patients.

A high- power ultrasonic method was used to prepare bilirubin oxidase nanoparticles (BOxNPs) which were immobilized on polyethylene (PE) film. The characterization of PE film bound to BOxNPs and BOxNPs was carried out using "Dynamic Light Scattering (DLS)," "Transmission Electron Microscopy (TEM)," and "Scanning Electron Microscopy (SEM)." The PE film was treated with nitric acid (HNO3) for its activation. BOxNPs bound to PE film exhibited optimal activity (pH-8), incubation time (11 s) with temperature 35 °C. A linear relationship was observed between the bilirubin concentrations (0.02-250 µM), with an apparent Km value and Vmax for PE- bound BOxNPs, at 0.015 µM and 2.56 µmol/mL/min. The mean recoveries of added serum bilirubin were 94.5 % at a level of 5 mM whereas 98.5 % were observed at 10 mM which showed the satisfactory reliability of BOxNPs immobilized on PE film. The coefficient of variation for serum bilirubin ranged between 4.52%-5.25%, measured on the first day (within batch) and after seven days of storage (between batch).This current method has showed a good correlation for bilirubin values when compared to the standard enzymatic colorimetric method using free enzyme. BOxNPs bound to PE film were reutilized 150 times with storage at 4 °C for 120 days.

Self-Assembled Two-Dimensional Molybdenum Disulfide Nanosheet Geno-Interface for the Detection of Salmonella.

This report presents a novel lab-on-a-paper (LoP)-based device coupled with a molybdenum disulfide nanosheet (MoS2NS)-modified electrochemical genosensor for detecting Salmonella-specific DNA. Conductive electrodes were grafted on a paper-based substrate employing a stencil printing technique, and MoS2NS was decorated on the working electrode. MoS2NS has strong affinity toward nucleo bases, which made it a best sensing interface for the immobilization of DNA. Morphological, optical, and structural characterizations were accomplished using X-ray diffraction (XRD), high-resolution transmission electron microscopy (HR-TEM), UV-vis spectroscopy (UV-vis), and Raman spectroscopy, repectively. The current studies of an electrochemical genosensor demonstrated a good linear detection range from 100-20 nM and a low limit of detection of 20 nM toward Salmonella DNA with R 2 = 0.991. The proposed LoP-based genosensor confirmed as a better sensing podium and an effectual immobilization matrix for DNA.

Determination of sulfite with emphasis on biosensing methods: a review.

Sulfite is used as a preservative in a variety of food and pharmaceutical industries to inhibit enzymatic and nonenzymatic browning and in brewing industries as an antibacterial and antioxidizing agent. Convenient and reproducible analytical methods employing sulfite oxidase are an attractive alternative to conventional detection methods. Sulfite biosensors are based on measurement of either O2 or electrons generated from splitting of H2O2 or heat released during oxidation of sulfite by immobilized sulfite oxidase. Sulfite biosensors can be grouped into 12 classes. They work optimally within 2 to 900 s, between pH 6.5 and 9.0, 25 and 40 °C, and in the range from 0 to 50,000 µM, with detection limit between 0.2 and 200 µM. Sulfite biosensors measure sulfite in food, beverages, and water and can be reused 100-300 times over a period of 1-240 days. The review presents the principles, merits, and demerits of various analytical methods for determination of sulfite, with special emphasis on sulfite biosensors.

An amperometric biosensor based on laccase immobilized onto Fe3O4NPs/cMWCNT/PANI/Au electrode for determination of phenolic content in tea leaves extract.

A method is described for the construction of an amperometric biosensor for detection of phenolic compounds based on covalent immobilization of laccase onto iron oxide nanoparticles (Fe3O4NPs) decorated carboxylated multiwalled carbon nanotubes (cMWCNTs)/polyaniline (PANI) composite electrodeposited onto a gold (Au) electrode. The modified electrode was characterized by scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The biosensor showed optimum response within 3s at pH 6.0 (0.1 M sodium acetate buffer) and 35°C, when operated at 0.3 V vs. Ag/AgCl. Linear range, detection limit were 0.1-10 µM (lower concentration range) and 10-500 µM (higher concentration range), and 0.03 µM respectively. The sensor measured total phenolic content in tea leaves extract. The enzyme electrode lost 25% of its initial activity after its 150 uses over a period of 4 months, when stored at 4°C.

Amperometric determination of total phenolic content in wine by laccase immobilized onto silver nanoparticles/zinc oxide nanoparticles modified gold electrode.

A method is described for construction of a highly sensitive amperometric biosensor for measurement of total phenolic compounds in wine by immobilizing laccase covalently onto nanocomposite of silver nanoparticles (AgNPs)/zinc oxide nanoparticles (ZnONPs) electrochemically deposited onto gold (Au) electrode. Scanning electron microscopy, X-ray diffraction, and electrochemical impedance spectroscopy were applied for characterization of the surface morphology of the modified electrode, and cyclic voltammetry was used to investigate the electrochemical properties of the proposed electrode toward the oxidation of guaiacol. The linearity between the oxidation current and the guaiacol concentration was obtained in a range of 0.1 to 500µM with a detection limit of 0.05µM (signal-to-noise ratio (S/N)=3) and sensitivity of 0.71µAµM(-1)cm(-2). The electrode showed increased oxidation and reduced reduction current with the deposition of AgNPs/ZnONPs on it. R(CT) values of ZnONPs/Au, AgNPs/ZnONPs/Au, and laccase/AgNPs/ZnONPs/Au electrode were 220, 175, and 380Ω, respectively. The biosensor showed an optimal response within 8s at pH 6.0 (0.1M acetate buffer) and 35°C when operated at 0.22V against Ag/AgCl. Analytical recovery of added guaiacol was 98%. The method showed a good correlation (r=0.99) with the standard spectrophotometric method, with the regression equation being y=1.0053x-3.5541. The biosensor lost 25% of its initial activity after 200 uses over 5months.

Development of an amperometric sulfite biosensor based on SO(x)/PBNPs/PPY modified ITO electrode.

A sulfite oxidase (SO(x)) (EC 1.8.3.1) purified from Syzygium cumini leaves was immobilized onto prussian blue nanoparticles/polypyrrole composite (PBNPs/PPY) electrodeposited onto the surface of indium tin oxide (ITO) electrode. An amperometric sulfite biosensor was fabricated using SO(x)/PBNPs/PPY/ITO electrode as working electrode, Ag/AgCl as standard and Pt wire as auxiliary electrode connected through a potentiostat. The working electrode was characterized by Fourier transform infrared (FTIR) spectroscopy, cyclic voltammetry (CV), scanning electron microscopy (SEM) and electrochemical impedance spectroscopy (EIS) before and after immobilization of SO(x). The biosensor showed optimum response within 2s, when operated at 20 mV s⁻¹ in 0.1M Tris-HCl buffer, pH 8.5 and at 35 °C. Linear range and minimum detection limit were 0.5-1000 µM and 0.12 µM (S/N=3) respectively. There was good correlation (r=0.99) between red wine samples sulfite value by standard DTNB method and the present method. The sensor was evaluated with 97% recovery of added sulfite in red wine samples and 2.2% and 4.3% within and between batch coefficients of variation respectively. The sensor was employed for determination of sulfite level in red and white wine samples. The enzyme electrode was used 200 times over a period of 3 months when stored at 4 °C.

Construction of amperometric uric acid biosensor based on uricase immobilized on PBNPs/cMWCNT/PANI/Au composite.

A chitosan-glutaraldehyde crosslinked uricase was immobilized onto Prussian blue nanoparticles (PBNPs) absorbed onto carboxylated multiwalled carbon nanotube (c-MWCNT) and polyaniline (PANI) layer, electrochemically deposited on the surface of Au electrode. The nanohybrid-uricase electrode was characterized by scanning electron microscopic (SEM), Fourier transform infrared spectroscopy (FTIR) and cyclic voltammetry. An amperometric uric acid biosensor was fabricated using uricase/c-MWCNT/PBNPs/Au electrode as working electrode, Ag/AgCl as standard and Pt wire as auxiliary electrode connected through a potentiostat. The biosensor showed optimum response within 4s at pH 7.5 and 40°C, when operated at 0.4V vs. Ag/AgCl. The linear working range for uric acid was 0.005-0.8 mM, with a detection limit of 5 µM. The sensor was evaluated with 96% recovery of added uric acid in sera and 4.6 and 5.4% within and between batch of coefficient of variation respectively and a good correlation (r=0.99) with standard enzymic colorimetric method. This sensor measured uric acid in real serum samples. The sensor lost only 37% of its initial activity after its 400 uses over a period of 7 months, when stored at 4°C.

An electrochemical sulfite biosensor based on gold coated magnetic nanoparticles modified gold electrode.

A sulfite oxidase (SO(X)) (EC 1.8.3.1) purified from Syzygium cumini leaves was immobilized onto carboxylated gold coated magnetic nanoparticles (Fe(3)O(4)@GNPs) electrodeposited onto the surface of a gold (Au) electrode through N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide (EDC)-N-hydroxy succinimide (NHS) chemistry. An amperometric sulfite biosensor was fabricated using SO(X)/Fe(3)O(4)@GNPs/Au electrode as working electrode, Ag/AgCl as standard and Pt wire as auxiliary electrode. The working electrode was characterized by Fourier Transform Infrared (FTIR) Spectroscopy, Cyclic Voltammetry (CV), Scanning Electron Microscopy (SEM) and Electrochemical Impedance Spectroscopy (EIS) before and after immobilization of SO(X). The biosensor showed optimum response within 2s when operated at 0.2V (vs. Ag/AgCl) in 0.1 M Tris-HCl buffer, pH 8.5 and at 35 °C. Linear range and detection limit were 0.50-1000 µM and 0.15 µM (S/N=3) respectively. Biosensor was evaluated with 96.46% recovery of added sulfite in red wine and 1.7% and 3.3% within and between batch coefficients of variation respectively. Biosensor measured sulfite level in red and white wines. There was good correlation (r=0.99) between red wines sulfite value by standard DTNB (5,5'-dithio-bis-(2-nitrobenzoic acid)) method and the present method. Enzyme electrode was used 300 times over a period of 4 months, when stored at 4 °C. Biosensor has advantages over earlier biosensors that it has excellent electrocatalysis towards sulfite, lower detection limit, higher storage stability and no interference by ascorbate, cysteine, fructose and ethanol.

An amperometric biosensor based on laccase immobilized onto MnO2NPs/cMWCNT/PANI modified Au electrode.

A method is described for construction of an amperometric biosensor for detection of phenolic compounds based on covalent immobilization of laccase (Lac) onto manganese dioxide nanoparticles (MnO(2)NPs) decorated carboxylated multiwalled carbon nanotubes (cMWCNTs)/PANI composite electrodeposited onto a gold (Au) electrode through N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxy succinimide (NHS) chemistry. The modified electrode was characterized by scanning electron microscopy (SEM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The biosensor showed optimum response at pH 5.5 (0.1M sodium acetate buffer) and 35°C, when operated at 0.3 V vs. Ag/AgCl. Linear range, response time, detection limit were 0.1-10 µM (lower concentration range) and 10-500 µM (higher concentration range), 4s and 0.04 µM, respectively. Biosensor measured total phenolic content in tea leaves extract. The enzyme electrode was used 150 times over a period of 5 months.

Development of an amperometric polyphenol biosensor based on fungal laccase immobilized on nitrocellulose membrane.

A method is described for construction of an amperometric polyphenol biosensor employing nitrocellulose membrane-bound laccase purified from cell-free extract of Ganoderma lucidum onto a Pt electrode. The biosensor showed optimum response within 10s, at 0.4 V in 0.1M acetate buffer, pH 6.0, and 35°C. Detection limit of the biosensor was 3.0 × 10(-8)M. Analytical recovery of added guaiacol was 97.00%. Within batch and between batch coefficients of variation were <0.97% and <1.26%, respectively. The sensor measured total phenolic content in fruit juices and alcoholic beverages. The enzyme electrode was used 100 times over 4 months, when stored at 4°C.

Development of an amperometric sulfite biosensor based on a gold nanoparticles/chitosan/multiwalled carbon nanotubes/polyaniline-modified gold electrode.

A sulfite oxidase (SOx) purified from leaves of Syzygium cumini (Jamun) was immobilized covalently onto a gold nanoparticles (AuNPs)/chitosan (CHIT)/carboxylated multiwalled carbon nanotubes (cMWCNTs)/polyaniline (PANI) composite that was electrodeposited onto the surface of a gold (Au) electrode. A novel and highly sensitive sulfite biosensor was developed that used this enzyme electrode (SOx/AuNPs/CHIT/cMWCNT/PANI/Au) as the working electrode, Ag/AgCl as the standard electrode, and Pt wire as the auxiliary electrode. The modified electrode was characterized by Fourier transform infrared (FTIR) spectroscopy, cyclic voltammetry (CV), scanning electron microscopy (SEM), and electrochemical impedance spectroscopy (EIS) before and after the immobilization of the SOx. The sensor produced its optimum response within 3 s when operated at 50 mVs(-1) in 0.1 M phosphate buffer, pH 7.0, and at 35 °C. The linear range and detection limit of the sensor were 0.75-400 µM and 0.5 µM (S/N = 3), respectively. The biosensor was employed to determine sulfite levels in fruit juices and alcoholic beverages. The enzyme electrode was used 300 times over a period of three months when stored at 4 °C.

Polyphenol biosensor based on laccase immobilized onto silver nanoparticles/multiwalled carbon nanotube/polyaniline gold electrode.

Laccase purified from Ganoderma sp. was immobilized covalently onto electrochemically deposited silver nanoparticles (AgNPs)/carboxylated multiwalled carbon nanotubes (cMWCNT)/polyaniline (PANI) layer on the surface of gold (Au) electrode. A polyphenol biosensor was fabricated using this enzyme electrode (laccase/AgNPs/cMWCNT/PANI/Au electrode) as the working electrode, Ag/AgCl as the reference electrode, and platinum (Pt) wire as the auxiliary electrode connected through a potentiostat. The biosensor showed optimal response at pH 5.5 (0.1 M acetate buffer) and 35°C when operated at a scan rate of 50 mV s(-1). Linear range, response time, and detection limit were 0.1-500 µM, 6 s, and 0.1 µM, respectively. The sensor was employed for the determination of total phenolic content in tea, alcoholic beverages, and pharmaceutical formulations. The enzyme electrode was used 200 times over a period of 4 months when stored at 4°C. The biosensor has an advantage over earlier enzyme sensors in that it has no leakage of enzyme during reuse and is unaffected by the external environment due to the protective PANI microenvironment.

Fabrication of polyphenol biosensor based on laccase immobilized on copper nanoparticles/chitosan/multiwalled carbon nanotubes/polyaniline-modified gold electrode.

A high-performance amperometric polyphenol biosensor was developed, based on covalent immobilization of Ganoderma sp. laccase onto copper nanoparticles (CuNP's)/chitosan (CHIT)/carboxylated multiwalled carbon nanotube (cMWCNT)/polyaniline (PANI)-modified gold (Au) electrode. The CuNP's and cMWCNT had a synergistic electrocatalytic effect in the matrix of CHIT. The biosensor showed optimum response at pH 6.0 (0.1 M acetate buffer) and 35°C, when operated at 50 mVs(-1). The biosensor exhibited excellent sensitivity (the detection limit was down to 0.156 µM for guaiacol), fast response time (less than 4s) and wide linear range (from 1 to 500 µM). Analytical recovery of added guaiacol was 96.40-98.46%. Within batch and between batch coefficients of variation were <2.6% and <5.3%, respectively. The enzyme electrode was used 300 times over a period of 7 months, when stored at 4°C.

An amperometric polyphenol biosensor based on laccase immobilized on epoxy resin membrane.

A polyphenol biosensor employing epoxy resin membrane bound Ganoderma sp. laccase has been developed. The biosensor showed optimum response within 30 s, when operated at 0.4 V in 0.1 M acetate buffer, pH 6.0 and 35 °C. Detection limit of the biosensor was 3.0 x 10-7 M. Analytical recovery of added guaiacol was 96.66%. Within batch and between batch coefficients of variation were <1.35% and <2.97% respectively. The biosensor was employed for amperometric determination of polyphenols in fruit juices and alcoholic beverages. The enzyme electrode was used 200 times over a period of 10 months, when stored at 4 °C.