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In the belief that DNA based
In the belief that DNA-based nanosensors and DNA microarrays should be easier to operate, faster, more accurate and more economically viable than PCR-based techniques, an electrochemical DNA biosensor was designed for detection of G. boninense[9], [10]. This was developed based on a gold electrode modified with a nanocomposite membrane of poly(3,4-ethylenedioxy thiophene)-poly(styrenesulfonate) containing gold nanoparticles. While the SM-406 showed successful detection of G. boninense, the preparation of representative samples and the extraction of DNA remained a challenge and invited further study.
Electrochemical biosensors work by producing an electrical signal that relates to the concentration of the biological analyte [11]. The relatively low cost and rapid response of DNA biosensors, in particular, promise exciting potential because of their simplicity, speed, and economical assays for gene analysis and testing.
In the construction of DNA biosensors, conducting polymers are effective platforms for the immobilization of biomolecules on electrode surfaces [12], providing good signal transduction, sensitivity, selectivity, durability, biocompatibility, direct electrochemical synthesis, and flexibility for the immobilization of DNA [13]. poly(3,4-ethyllenedioxythiophen)–poly(styrenesulfonate) (PEDOT-PSS) has shown good potential due to its function (Ф) ∼5 eV [14]. It needs to be combined with poly(styrenesulfonate) (PSS) to enable its dispersion in water for forming thin films on surfaces [15].
Additionally, the explosion of nanotechnology and the use of nanomaterials in DNA biosensors is further enhancing immobilization and interface capabilities of the DNA material with the electrode and, ultimately, the detection signal response [16], [17]. Metal nanoparticles are frequently added to conjugate polymers to boost the conductivity of the surface [15], [18], [19] and to increase the active surface area of the sensor, ultimately enhancing the transduction signal response [20], [21], [22]. In this work, gold (Au) was used.
Magnetic nanoparticles (MNPs), particularly iron oxide Fe3O4, are especially popular because of their powerful magnetic properties, large surface areas and the ease in which they can be separated from a liquid with a magnet [23].
Materials and methods
Results and discussion
Conclusion
Electrochemical nanosensors are important analytical tools as the demand for sensitive, rapid, and selective detection of analytes increases in the fields of healthcare, environmental monitoring, and biological analysis and are already demonstrating broad application and some early success as reliable, portable devices [37].
However, researchers are still grappling with designing increased specificity and sensitivity as well as resolving a range of other issues including biocompatibility and stability [37].
Building on the earlier effort, we continued to use a modified gold electrode and gold nanoparticles but this time we attached functionalised Fe3O4 magnetic nanoparticles to the Capture probe. This enabled a form of indirect sensing and successfully facilitated bringing a representative sequence of the analyte to the biorecognition surface.
Whilst the challenge of detecting small Target sequences in large amounts of extracted genomic DNA is not yet over in the search for early detection of Ganoderma, it is hoped that the technique described here can make a positive contribution and will be quickly developed further not only in the detection of a wide range of pathogens but also any types of genomic DNA.
Conflict of interest
Acknowledgments
This research was supported by the Malaysian Ministry of Science, Technology and Innovation. The authors would also like to thank Jill Jamieson for editorial assistance.
Introduction
Anthracyclines, as exemplified by daunorubicin (DNR), are a class of anti-neoplastic agents widely used for the treatment of malignancy. Their cytotoxic mechanism involves the production of DNA damage through intercalating with DNA and the inhibition of topoisomerase (topo) II, finally causing DNA-double-strand breaks (DSBs) and inducing apoptosis [1]. Drug resistance is a major obstacle in the successful treatment of leukemia and solid tumors and is still a major cause of death in leukemia and cancer patients [2]. The development of a resistance mechanism in response to doxorubicin-induced apoptosis includes P-glycoprotein and Bcl-2 overexpression, altered topo II activity, and loss of p53 function, etc. [3]. For example, overexpression of P-glycoprotein confers resistance to a variety of structurally and functionally unrelated anti-cancer drugs, a function known as multidrug resistance (MDR).