Voltammetric determination of human papillomavirus 16 DNA by using interdigitated electrodes modified with titanium dioxide nanoparticles ABSTRACT A gene sensor for rapid detection of the Human Papillomavirus 16 (HPV 16) which is associated with the appearance of cervical cancer was developed. The assay is based on voltammetric determination of HPV 16 DNA by using interdigitated electrodes modified with titanium dioxide nanoparticles. Titanium dioxide nanoparticles (NPs) were used to modify a semiconductor-based interdigitated electrode (IDE). The surface of the NPs was then functionalized with a commercial 24-mer oligomer DNA probe for HPV 16 that was modified at the 5′ end with a carboxyl group. If the probe interacts with the HPV 16 ssDNA, the current, best measured at a working voltage of 1.0 V, increases. The gene sensor has has a ∼ 0.1 fM limit of detection which is comparable to other sensors. The dielectric voltammetry analysis was carried out from 0 V to 1 V. The electrochemical sensitivity of the IDE is 2.5 × 10⁻⁵ μA•μM⁻¹•cm⁻². Graphical abstract Schematic of an interdigitated electrode (IDE) modified with titanium dioxide nanoparticles for voltammetric determination of HPV 16 DNA by using an appropriate DNA probe.
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Spontaneous unexplained preterm labor with intact membrane (sPTL-IM) remains as an unresolved challenge in obstetrics due to the complex syndromes involved during preterm birth. Two dimensional-gel electrophoresis (2D-GE) coupled with matrixassisted laser desorption/ionization-time of flight/time of flight (MALDI TOF/TOF) mass spectrometry has become an alternative in screening for potential novel protein-based biomarkers and revealing the pathophysiology of sPTL-IM. To achieve this objective, protein extracted from fetal and maternal sides of the placenta obtained from sPTL-IM (n = 5) and the respective control (n = 10) groups were separated and compared using 2D-gel electrophoresis. MALDI-TOF/TOF mass spectrometry was utilized to identify the differentially expressed proteins between both groups, and the molecular functions of these proteins were studied. A total of 12 proteins were significantly differentiated in sPTL-IM over the control. Differentially expressed proteins were identified to have involved in structural/cytoskeletal components, immune responses, fetal and placenta development, and anticoagulation cascade. More proteins were found to be differentially expressed in the fetal side compared to the maternal side of the placenta. This postulates that the influence of sPTL-IM from fetus is greater than that of the mother. Ultimately, these results might lead to further investigations in elucidating the potential of these proteins as biomarkers and/or drug targets.
Objective To evaluate the association between transvaginal ultrasound scan of cervix and Bishop’s score in predicting successful induction of labour, cut-off points and patients’ tolerability and acceptance for both procedures. Design A comparative clinical trial. Setting A tertiary hospital in Selangor, Malaysia. Participants 294 women planned for elective induction of labour for various indications were included. All women had transvaginal ultrasound to assess the cervical length and digital vaginal examination to assess the Bishop cervical scoring by separate investigators before induction of labour. Primary outcome measure To evaluate the association of the cervical length by transvaginal ultrasound scan and Bishop score in predicting successful induction of labour. Secondary outcome measure Variables associated with successful induction of labour and patients’ tolerability and acceptance for transvaginal ultrasound scan of cervix. Results There was no statistically significant difference among the vaginal and Caesarean delivery groups in terms of mean maternal age, height, weight, body mass index, ethnicity and gestational age at induction. Vaginal delivery occurred in 207 women (70.4%) and 87 women (29.6%) delivered via Caesarean section. There was a high degree of correlation between the cervical length and Bishop score (r-value 0.745; p <0.001). Sonographic assessment of cervical length demonstrated a comparable accuracy in comparison to Bishop score. Analysis using ROC curves noted an optimal cut-off value of ≤27mm for cervical length and Bishop score of ≥ 4, with a sensitivity of 69.1% vs 67%, specificity 60.9% vs 55%, and area under the curves (AUCs) of 0.672 and 0.643 respectively (p <0.001). Multivariate logistic regression analysis demonstrated that parity (OR 2.70), cervical length (OR 0.925), Bishop score (OR 1.272) and presence of funnelling (OR 3.292) were highly significant as independent predictors of success labour induction. Women also expressed significantly less discomfort with transvaginal ultrasound compared with digital vaginal examination. Conclusion Sonographic assessment of cervical measurement predicts the success of induction of labour with similar diagnostic accuracy with conventional Bishop score.
The quest for alternative methods is driven by the need to provide expertise in real time in biological fields such as medicine, pathogenic bacteria and viruses identification, food protection, and quality control. Polymerase Chain Reaction (PCR) and Enzyme Linked Immunosorbent Assay (ELISA) are examples of traditional methods that have some limitations and lengthy procedures. Biosensors are the most appealing option because they provide easy, dependable, fast, and selective detection systems compared to conventional methods. This review provides an overview of electrochemical genosensor based biosensor diagnostic system for infectious diseases detection as well as their applications, demonstrating their utility as a fast and responsive tool for detecting pathogenic bacteria, viruses, GMOs, and human diseases.
Effective protein extraction is essential especially in producing a well-resolved proteome on 2D gels. A well-resolved placental cotyledon proteome, with good reproducibility, have allowed researchers to study the proteins underlying the physiology and pathophysiology of pregnancy. The aim of this study is to determine the best protein extraction protocol for the extraction of protein from placental cotyledons tissues for a two-dimensional gel electrophoresis (2D-GE). Based on widely used protein extraction strategies, 12 different extraction methodologies were carefully selected, which included one chemical extraction, two mechanical extraction coupled protein precipitations, and nine chemical extraction coupled protein precipitations. Extracted proteins were resolved in a one-dimensional gel electrophoresis and 2D-GE; then, it was compared with set criteria: extraction efficacy, protein resolution, reproducibility, and recovery efficiency. Our results revealed that a better profile was obtained by chemical extraction in comparison to mechanical extraction. We further compared chemical extraction coupled protein precipitation methodologies, where the DNase/lithium chloride-dense sucrose homogenization coupled dichloromethane-methanol precipitation (DNase/LiCl-DSH-D/MPE) method showed good protein extraction efficiency. This, however, was carried out with the best protein resolution and proteome reproducibility on 2D-gels. DNase/LiCl-DSH-D/MPE was efficient in the extraction of proteins from placental cotyledons tissues. In addition, this methodology could hypothetically allow the protein extraction of any tissue that contains highly abundant lipid and glycogen.
The aim of this work was to develop a novel, simple, inexpensive, sensitive an electrochemical DNA biosensor based on interdigitated electrodes (IDEs) integrated gold nanoparticle modified electrode by electro less deposition for HPV 18. The biosensor was designed with a 30 mer E6 region of HPV 18 DNA modified probe. The E6 region has been used for their clinical importance properties and suitable as recognition biomarker region. Three different target types were tested which complementary target, non-complementary target and mismatch target. All target were analyzed for detection of HPV 18 in early stages by using Dielectric Analyzer (DA), Alpha-A High-performance Frequency Analyzer, Novocontrol Technologies, Handsagen, Germany associated with the software package Windeta. Complementary target gives a positive result in HPV detection, while non-complementary and mismatch target give negative results. IDE device with 5 nm gap sizes has demonstrated a high performance towards the detection of HPV18 ssDNA target by modified with 30 nm gold nanoparticle. The electrochemical biosensor showed better performance compared to agarose gel electrophoresis assay. This technology can be used as a new and attractive sensor development for detection of virus infection in human bodies.
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