A novel interdigitated capacitor based biosensor for detection of cardiovascular risk marker

C-reactive protein (CRP) is a potential biomarker whose elevated levels in humans determine cardiovascular disease risk and inflammation. In this study, we have developed a novel capacitive biosensor for detection of CRP-antigen using capacitor with interdigitated gold (GID) electrodes on nanocrystalline diamond (NCD) surface. The NCD surface served as a dielectric layer between the gold electrodes. GID-surface was functionalized by antibodies and the immobilization was confirmed by Fourier transform spectroscopy (FT-IR) and contact angle measurements. The CRP-antigen detection was performed by capacitive/dielectric-constant measurements. The relaxation time and polarizability constants were estimated using Cole-Cole model. Our results showed that the relaxation time constant (tau) of only CRP-antibody was within 10(-16)-10(-13) s, which was increased to 10(-11) s after the incubation with CRP-antigen, suggesting that the CRP-antigen was captured by the antibodies on GID-surface. In addition, polarizability constant(m) of CRP was also increased upon incubation with increasing concentration of CRP-antigen. Our results showed that the response of GID-NCD-based capacitive biosensor for CRP-antigen was dependent on both concentration (25-800 ng/ml) as well as frequency (50-350 MHz). Furthermore, using optimized conditions, the GID-NCD based capacitive biosensor developed in this study can potentially be used for detection of elevated levels of protein risk markers in suspected subjects for early diagnosis of disease

E. coli-quantum dot bioconjugates as whole-cell fluorescent reporters for probing cellular damage

A quantum dot (QD) conjugated whole-cell E. coli biosensor (E. coli–QD bioconjugates) was developed as a new molecular tool for probing cellular damage. The E. coli–QD bioconjugates were viable and exhibited fluorescence emission at 585 nm. Scanning electron microscopy (SEM) analysis of E. coli–QD bioconjugates revealed that the QDs were immobilized on the cell-surfaces and the fluorescence emission from QDs present on cell-surfaces was visualized by confocal microscopic examination. The E. coli–QD bioconjugates were employed as whole-cell fluorescent reporters that were designed to function as fluorescence switches that turn-off when cellular damage occurs. In this study, multi-walled carbon nanotubes (CNTs) were utilized as a model nanomaterial to probe cellular damage. Fluorescence spectra were recorded after the exposure of E. coli–QD bioconjugates with CNTs. We observed a strong correlation between fluorescence emission spectra, SEM and confocal microscopic analysis demonstrating that CNTs induced a dose and exposure time-dependent cellular toxicity. This toxicity mainly occurred by the physical interaction and cellular trafficking mechanisms that led to the collapse of the cellular structure and thus loss of fluorescence. The responses of E. coli–QD bioconjugates against CNTs were also visualized by simply exposing the cells to UV light and therefore rapid toxicity analysis and screening can be made. Our study demonstrated an easy and simple method to determine an important mechanistic perspective for the biological toxicity of chemicals or nanomaterials (NMs)

Biosensors for cardiac biomarkers detection: a review

The cardiovascular disease (CVD) is considered as a major threat to global health. Therefore, there is a growing demand for a range of portable, rapid and low cost biosensing devices for the detection of CVD. Biosensors can play an important role in the early diagnosis of CVD without having to rely on hospital visits where expensive and time-consuming laboratory tests are recommended. Over the last decade, many biosensors have been developed to detect a wide range of cardiac marker to reduce the costs for healthcare. One of the major challenges is to find a way of predicting the risk that an individual can suffer from CVD. There has been considerable interest in finding diagnostic and prognostic biomarkers that can be detected in blood and predict CVD risk. Of these, C-reactive protein (CRP) is the best known biomarker followed by cardiac troponin I or T (cTnI/T), myoglobin, lipoprotein-associated phospholipase A(2), interlukin-6 (IL-6), interlukin-1 (IL-1), low-density lipoprotein (LDL), myeloperoxidase (MPO) and tumor necrosis factor alpha (TNF-α) has been used to predict cardiovascular events. This review provides an overview of the available biosensor platforms for the detection of various CVD markers and considerations of future prospects for the technology are addressed

Review on carbon-derived, solid-state, micro and nano sensors for electrochemical sensing applications

The aim of this review is to summarize the most relevant contributions in the development of electrochemical sensors based on carbon materials in the recent years. There have been increasing numbers of reports on the first application of carbon derived materials for the preparation of an electrochemical sensor. These include carbon nanotubes, diamond like carbon films and diamond film-based sensors demonstrating that the particular structure of these carbon material and their unique properties make them a very attractive material for the design of electrochemical biosensors and gas sensors. Carbon nanotubes (CNT) have become one of the most extensively studied nanostructures because of their unique properties. CNT can enhance the electrochemical reactivity of important biomolecules and can promote the electron-transfer reactions of proteins (including those where the redox center is embedded deep within the glycoprotein shell). In addition to enhanced electrochemical reactivity, CNT-modified electrodes have been shown useful to be coated with biomolecules (e.g., nucleic acids) and to alleviate surface fouling effects (such as those involved in the NADH oxidation process). The remarkable sensitivity of CNT conductivity with the surface adsorbates permits the use of CNT as highly sensitive nanoscale sensors. These properties make CNT extremely attractive for a wide range of electrochemical sensors ranging from amperometric enzyme electrodes to DNA hybridization biosensors. Recently, a CNT sensor based fast diagnosis method using non-treated blood assay has been developed for specific detection of hepatitis B virus (HBV) (human liver diseases, such as chronic hepatitis, cirrhosis, and hepatocellular carcinoma caused by hepatitis B virus). The linear detection limits for HBV plasma is in the range 0.5–3.0 μL−1 and for anti- HBVs 0.035–0.242 mg/mL in a 0.1 M NH4H2PO4 electrolyte solution. These detection limits enables early detection of HBV infection in suspected serum samples. Therefore, non-treated blood serum can be directly applied for real-time sensitive detection in medical diagnosis as well as in direct in vivo monitoring. Synthetic diamond has been recognized as an extremely attractive material for both (bio-) chemical sensing and as an interface to biological systems. Synthetic diamond have outstanding electrochemical properties, superior chemical inertness and biocompatibility. Recent advances in the synthesis of highly conducting nanocrystalline-diamond thin films and nano wires have lead to an entirely new class of electrochemical biosensors and bio-inorganic interfaces. In addition, it also combines with development of new chemical approaches to covalently attach biomolecules on the diamond surface also contributed to the advancement of diamond-based biosensors. The feasibility of a capacitive field-effect EDIS (electrolyte-diamond-insulatorsemiconductor) platform for multi-parameter sensing is demonstrated with an O-terminated nanocrystalline-diamond (NCD) film as transducer material for the detection of pH and penicillin concentration. This has also been extended for the label-free electrical monitoring of adsorption and binding of charged macromolecules. One more recent study demonstrated a novel bio-sensing platform, which is introduced by combination of a) geometrically controlled DNA bonding using vertically aligned diamond nano-wires and b) the superior electrochemical sensing properties of diamond as transducer material. Diamond nanowires can be a new approach towards next generation electrochemical gene sensor platforms. This review highlights the advantages of these carbon materials to promote different electron transfer reactions specially those related to biomolecules. Different strategies have been applied for constructing carbon material-based electrochemical sensors, their analytical performance and future prospects are discussed

Nanocrystalline diamond film for biosensor applications

In this study, we have developed a novel capacitive biosensor based oil interdigitated gold nanodiamond (GID-NCD) electrode for detection of C-reactive protein (CRP) antigen. CRP is one of the plasma proteins known as acute-phase proteins and its levels rise dramatically during inflammatory processes occurring in the body. It has been reported that CRP in serum can be used for risk assessment of cardiovascular diseases. The antibodies immobilization were confirmed by Fourier transform spectroscopy (FTIR) and contact angle measurements. In this capacitive biosensor, nanocrystalline diamond acting as a dielectric layer between the electrodes. The CRP antigen detection was performed by capacitive/dielectric-constant measurements. Our results showed that the response of NCD-based capacitive-based biosensor for CRP antigen was dependent on both concentration (25-800 ng/ml) as well as frequency (50-350 MHz). Furthermore, using optimized conditions, the biosensors developed in this study can be potentially used for detection of elevated level of risk markers protein in suspected subjects for early diagnosis of disease

Toxicity evaluation of e-juice and its soluble aerosols generated by electronic cigarettes using recombinant bioluminescent bacteria responsive to specific cellular damages

Electronic-cigarettes (e-cigarette) are widely used as an alternative to traditional cigarettes but their safety is not well established. Herein, we demonstrate and validate an analytical method to discriminate the deleterious effects of e-cigarette refills (e-juice) and soluble e-juice aerosol (SEA) by employing stress-specific bioluminescent recombinant bacterial cells (RBCs) as whole-cell biosensors. These RBCs carry luxCDABE-operon tightly controlled by promoters that specifically induced to DNA damage (recA), superoxide radicals (sodA), heavy metals (copA) and membrane damage (oprF). The responses of the RBCs following exposure to various concentrations of e-juice/SEA was recorded in real-time that showed dose-dependent stress specific-responses against both the e-juice and vaporized e-juice aerosols produced by the e-cigarette. We also established that high doses of e-juice (4-folds diluted) lead to cell death by repressing the cellular machinery responsible for repairing DNA-damage, superoxide toxicity, ion homeostasis and membrane damage. SEA also caused the cellular damages but the cells showed enhanced bioluminescence expression without significant growth inhibition, indicating that the cells activated their global defense system to repair these damages. DNA fragmentation assay also revealed the disintegration of total cellular DNA at sub-toxic doses of e-juice. Despite their state of matter, the e-juice and its aerosols induce cytotoxicity and alter normal cellular functions, respectively that raises concerns on use of e-cigarettes as alternative to traditional cigarette. The ability of RBCs in detecting both harmful effects and toxicity mechanisms provided a fundamental understanding of biological response to e-juice and aerosols.open

Determination of Reference Range of Glycosylated Haemoglobin (HbA1c) for Different Age Groups

Objective: To determine the reference interval of glycosylated haemoglobin for different age groups in our local Pakistani population. Subjects & methods: Total number of 164 healthy adults (91 females and 73 males), were selected through nonprobability consecutive sampling. After informed consent, venous blood sample (3-4 ml) from each subject was collected in EDTA vial under aseptic conditions. Haemolysed blood was used for estimation of HbA1c by Fast Ion-Exchange Resin Separation method on Microlab 300. The participants of the study were divided into three groups according to their age distribution. All data was analysed using Microsoft Excel and statistical software Minitab15. Mean and SDs were calculated for each group. Reference range of HbA1c was calculated for each group by using the formula Mean ± 2SD. Results: One hundred and sixty-four healthy individuals were included in the study. Mean age of the individuals was 32±11.6 years and mean HbA1c level was 4.17±0.93%. The reference range of HbA1c for age group 20-30 years was 2.09-5.57% with the mean value of 3.83±0.87%. The reference range of HbA1c for age group 31-40 years was 2.63-5.99% with the mean value of 4.31±0.84%.The reference range of HbA1c for individuals > 40 years of age was 2.8-6.4% with the mean value of 4.31±0.9%. Conclusion: For proper assessment of the diabetic status, it is mandatory that every lab should establish its own reference range of HbA1c for both genders and different age groups representing its patient population.&nbsp