Ongoing ResearchÂ
Design and fabrication of the electronic test card.
Development of protocol for RNA detection.
Validation of the card on actual clinical samples.
The present research proposal will focus on development of a low-cost electronic test card for screening the presence of Viral RNA in clinical samples. We would amplify the RNA through one step Reverse transcription loop-mediated isothermal amplification (RT- LAMP) process. Simple, low cost, point of care diagnosis is the need for developing countries like India and LAMP is an ultrasensitive nucleic acid amplification method that can detect minute quantities of DNA or RNA templates within roughly an hour without the requirement of any specialized or skilled technicians. LAMP PCR and RT-PCR is a simple and sensitive method which finds wide application in detection of infectious diseases, which is usually done using visual colorimetric detection. Here, we are proposing a highly integrable and fast electronic detection scheme which costs quite lesser than the colorimetric one. The specificity in the detection would be included by integrating gold nanoparticle labelling (inexpensive as compared to electronic/electrochemical labels) to the amplifying sequence and specifically capturing it through di-electrophoresis (DEP). The more optimized electrode platform on silicon would be realized to carry out the stated process. The stated scheme would be rather simpler as it would need only a single temperature window instead of the variable temperature sequences in PCR. The validation will be done by using HCV RNA detection in blood samples and comparing it with Real Time PCR. Therefore, in terms of applications of the proposed scheme, the current industrial application is for the detection of Hepatitis C virus RNA (HCVRNA).
Figure. Illustration of the proposed area of research.
The above point of care test will help in reaching out to many and will increase manifolds the uptake of testing, identification of patients who will be treated and therefore helping in achievement of elimination of HCV. This witnesses the immense requirement of RNA detection in Hepatitis diagnostics.
