R428 Supplier The main objective of this research was to dev

The main objective of this research was to develop a simple and fast method to create a well-defined recognition surface for the immobilization and hybridization of the Trichoderma harzianum gene. The incorporation of ILs, ZnO nanoparticles and CHIT nanocomposite membrane were explored to increase the electrochemical signals of the redox indicator and to enhance the sensitivity for DNA detection. The analytical performance of the designed electrochemical biosensor was evaluated for the detection of a specific sequence related to a T. harzianum gene based on the internal transcribed spacer 1 and 2 regions of the rDNA. The developed DNA biosensor was also applied in the analysis of crude DNA fragments.
Materials and methods
Results and discussion
Conclusion An electrochemical DNA biosensor is successfully developed based on ILs/ZnO nanoparticles/CHIT/AuE then applied for the detection of DNA immobilization and hybridization from T. harzianum gene and crude DNA taken from real samples. ZnO nanoparticles homogeneously dispersed in CHIT and ILs film enhanced the consignment interface for the immobilization of probe DNA and increased the detection sensitivity level for DNA hybridization. This developed DNA biosensor was applied in the detection of T. harzianum gene sequences in a wide concentration range of 1.0×10−18–1.82×10−4molL−1 (n=5) using DPV method and methylene blue used as an electrochemical indicator. The designed biosensor has several advantages such as a simple preparation procedure, high selectivity, low cost, fast response, and a wide linear range. This developed biosensor also offers a convenient research tool for the identification of T. harzianum in R428 Supplier to non-harzianum.
Conflict of interest
Acknowledgment The authors would like to thank the Ministry of Science, Technology and Innovation, Malaysia (MOSTI) for their support under research project no. SCF0090-IND-2013.
Introduction Modern living is overwhelmed by the ever increasing costs of treatment and care for non-healing chronic wounds such as venous leg ulcers, diabetic foot ulcers etc. [1]. Australia alone spends almost $2.5 billion annually on wound management with about 4000 limb amputations per year [2]. Chronic wound management also creates a burden in terms of nursing care and hospitalization time. During 2010, almost 6.5% of total diseases and 31% of nursing activities were attributed to wound management in Australia [3]. The USA has spent almost $3 billion during 2006–07 [4], while the UK has spent £26 million during 2008–09 [1] for the treatment of non-healing chronic wounds. Throughout the world, this cost is on the rise with an estimate of over $20 billion in the near future [5]. Understanding the healing trajectory of wounds is important to mitigate the effects of non-healing wounds on society and on the economy. Wound healing is a complex biochemical process divided into four distinct phases (i) haemostasis, (ii) inflammation, (iii) proliferation, and (iv) remodelling [4]. A wound is properly healed up if all of these phases occur in their natural sequence. However, if one or more stages are prolonged for any reason, the healing process is impaired, resulting in delayed or non-healing wounds [6], [7]. Many factors contribute to impaired healing, including oxygenation, age, gender, infection, diet, hormones, stress, diabetes, medications etc. [4]. Covering a chronic wound with suitable dressing is the most economical and effective way of treatment [8]. For some wounds, appropriate pressure bandages are also applied to absorb the wound exudate and expedite healing [9]. Healing rate may be increased by providing external moisture to the wound site through moisture-retentive dressings [10]. When healing progresses, wound exudate gradually diminishes. At this stage, a special type of dressing is required (e.g. foams, hydrogels or hydrocolloids) to maintain the required moisture level at the wound site. Otherwise, the wound may not heal due to hypoxia [11]. Furthermore, wound-site temperature and pH level may also be used with other parameters, such as moisture level, nitric oxide concentration etc., to predict the status of wound healing [12], [13], and hence the proper time to change the dressings may be determined. However, in current clinical practice, most of the dressings are changed routinely without any scientific reason or clinical evidence which complicates the problem of impaired wound healing.