Exploring the feasibility of developing a rapid diagnostic test kit against Dengue virus using nanotechnology
Postgraduate student:
Ms. M.M. Erandi Munasinghe, Molecular Medicine Unit, Faculty of Medicine, University of Kelaniya, Ragama, Sri Lanka.
Supervisors:
Prof. W. Abeyewickreme - Dept. of Parasitology and Molecular Medicine Unit, Faculty of Medicine, University of Kelaniya.
Dr. P.A.D.H.N. Gunathilaka - Biotechnology Unit, Industrial Technology Institute, Colombo 7.
Dr. A.M.M.H. Athapaththu - Biotechnology Unit, Industrial Technology Institute, Colombo 7.
Dr. P.A.D.H.N. Gunathilaka - Biotechnology Unit, Industrial Technology Institute, Colombo 7.
Dr. A.M.M.H. Athapaththu - Biotechnology Unit, Industrial Technology Institute, Colombo 7.
Objectives
General Objectives
General Objectives
- To develop a diagnostic kit using polyclonal/monoclonal antibodies to detect Dengue virus from blood at an early stage Specific Objectives
- To develop polyclonal/monoclonal antibodies against NS1 region of DENV
- To determine the feasibility of a nanomaterial to anchor the developed antibodies
Project Summary
Early diagnosis of dengue has become a major in clinical setup. Diagnosis of the disease as early as possible would improve the patient management, vector controlling and lower the fatality rate. During the past decade there have been major advances and breakthroughs in dengue diagnosis with methods such as Polymerase Chain Reaction (PCR), Real Time (RT) PCR, Enzyme Linked Immunosorbant Assay (ELISA) and recombinant protein assays. Even though current available methods are sensitive, confirmation of the disease at the day one of onset of illness is questionable. Further, these advanced methods lack the detection of virus at single cell level. Most of the hospitals use antibody detection through ELISA, which is too late for patient management. Therefore there is an urgent need to develop a diagnostic method that could be used for confirmation of the virus at the earliest. Further there is a need to develop a kit that could be used easily in field settings such as home or rural clinics where no facilities needed. Nanoparticles have displayed remarkable potential for detecting pre-cancerous cells, disease markers and fragments of viruses at single cell level (Mohamad et al., 2012). Most importantly, with the modifications of the nanostructures' surface, a single binding event can be potentially recorded leading to the detection at single cell level. This would provide proper patients care ultimately preventing unnecessary deaths and lowering the government expenditure for numerous unnecessary diagnostic tests. Early detection of the disease could be treated quickly and make full recoveries.
In less than a decade, biosensors based on nanotechnology have successfully made the transition from proof of concept to highly selective, ultra-sensitive devices capable of detecting specific proteins and DNA sequences. These devices utilize a capture agent on the sensor surface to selectively bind the target biomolecules. The captured biomolecules affect the electronic properties of the nanowires/nanotubes, resulting in an electronically readable signal. Capture agents commonly used in Nano biosensors include antibodies, oligonucleotides, and small ligands (e.g. biotin) (5). Therefore, combination of biotechnology and nanotechnology is ideal for producing highly sensitive and specific small portable test strips which could be bought over the counter and easily discarded once used.
In this study a portable, user friendly and cheap kit that could be used as a home appliance or in any field setting for detection of the DENV in blood at single cell level would be developed by combining biotechnology and nanotechnology. Therefore, the intension is to determine the feasibility of developing a rapid diagnostic kit to an extent that public could purchase the test strips over the counter from any pharmacy. The study will also focus on developing antibodies against DENV, formulating a nanomaterial which can anchor those antibodies and thereby to facilitate with highly sensitive, portable and cheap diagnostic kit to detect dengue virus from blood, at early viremic phase. Finally, the results of the current research will lead to a nanotechnology-based kit for the detection of DENV thereby to prevent dengue epidemics through early diagnosis, proper treatment and patient management in order to safeguard human and economic wellness.
Early diagnosis of dengue has become a major in clinical setup. Diagnosis of the disease as early as possible would improve the patient management, vector controlling and lower the fatality rate. During the past decade there have been major advances and breakthroughs in dengue diagnosis with methods such as Polymerase Chain Reaction (PCR), Real Time (RT) PCR, Enzyme Linked Immunosorbant Assay (ELISA) and recombinant protein assays. Even though current available methods are sensitive, confirmation of the disease at the day one of onset of illness is questionable. Further, these advanced methods lack the detection of virus at single cell level. Most of the hospitals use antibody detection through ELISA, which is too late for patient management. Therefore there is an urgent need to develop a diagnostic method that could be used for confirmation of the virus at the earliest. Further there is a need to develop a kit that could be used easily in field settings such as home or rural clinics where no facilities needed. Nanoparticles have displayed remarkable potential for detecting pre-cancerous cells, disease markers and fragments of viruses at single cell level (Mohamad et al., 2012). Most importantly, with the modifications of the nanostructures' surface, a single binding event can be potentially recorded leading to the detection at single cell level. This would provide proper patients care ultimately preventing unnecessary deaths and lowering the government expenditure for numerous unnecessary diagnostic tests. Early detection of the disease could be treated quickly and make full recoveries.
In less than a decade, biosensors based on nanotechnology have successfully made the transition from proof of concept to highly selective, ultra-sensitive devices capable of detecting specific proteins and DNA sequences. These devices utilize a capture agent on the sensor surface to selectively bind the target biomolecules. The captured biomolecules affect the electronic properties of the nanowires/nanotubes, resulting in an electronically readable signal. Capture agents commonly used in Nano biosensors include antibodies, oligonucleotides, and small ligands (e.g. biotin) (5). Therefore, combination of biotechnology and nanotechnology is ideal for producing highly sensitive and specific small portable test strips which could be bought over the counter and easily discarded once used.
In this study a portable, user friendly and cheap kit that could be used as a home appliance or in any field setting for detection of the DENV in blood at single cell level would be developed by combining biotechnology and nanotechnology. Therefore, the intension is to determine the feasibility of developing a rapid diagnostic kit to an extent that public could purchase the test strips over the counter from any pharmacy. The study will also focus on developing antibodies against DENV, formulating a nanomaterial which can anchor those antibodies and thereby to facilitate with highly sensitive, portable and cheap diagnostic kit to detect dengue virus from blood, at early viremic phase. Finally, the results of the current research will lead to a nanotechnology-based kit for the detection of DENV thereby to prevent dengue epidemics through early diagnosis, proper treatment and patient management in order to safeguard human and economic wellness.