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16 result(s) for "Gorthi, Sai Siva"
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Rapid diagnosis of Plasmodium falciparum malaria using a point-of-care loop-mediated isothermal amplification device
LAMP diagnosis of malaria is simple and cost-effective with acceptable sensitivity and specificity as compared to standard diagnostic modules such as microscopy, RDTs and nested PCR, and thus its deployment for onsite screening of malaria in resource-limited regions is under consideration. However, the requirement of an electricity-operated dry bath and bulky read-out unit is still a major concern. In an effort to simplify this limitation, we have developed a portable LAMP device and fluorescence readout unit which can be used in the rapid point-of-care diagnosis of malaria. We have developed a point-of-care diagnostic LAMP device that is easy to operate by a mobile application, and the results can be quantified with a fluorescent readout unit. The diagnostic performance of the device was evaluated in 90 P . falciparum -infected clinical isolates stored at 4°C for 6-7 years and 10 freshly collected isolates from healthy volunteers. The LOD and quantitative ability of LAMP in estimating parasitemia levels were revealed with laboratory-grown P. falciparum strain (3D7). The LAMP assay performed in our device was exclusive for P. falciparum detection with sensitivity and specificity determined to be 98.89% and 100%, respectively, in clinical isolates. The LOD was documented to be 1 parasite/µl at the cut-off ADC value of 20. Parasite density estimated from ADC values showed concordance with microscopically determined parasite density of the cultured P. falciparum 3D7 strain. The LAMP assay performed in our device provides a possible portable platform for its deployment in the point-of-care diagnosis of malaria. Further validation of the quantitative ability of the assay with freshly collected or properly stored clinical samples of known parasitemia is necessary for field applicability.
Hand-Powered Elastomeric Pump for Microfluidic Point-of-Care Diagnostics
The pumping of fluids into microfluidic channels has become almost an unavoidable operation in all microfluidic applications. Such a need has seen an outburst of several techniques for pumping, out of which the majority of techniques involve complicated fabrication, as they require the introduction of electrodes, valves, piezoelectric materials, acoustic transducers, etc., into the microfluidic device. In addition to the complexity, this also escalates the cost incurred per device. Further, the use of stable external power supplies to produce such a pumping action adds to the bulkiness of the pumps, making them unsuitable for point-of-care diagnostic (POCD) applications. This paper reports a technique of pumping that is simple to realize and does not require external electric/magnetic power, but exploits the elastic properties of materials to achieve the pumping action. This mechanism of pumping ensured the cost per pump to less than 4 USD and can be used for at least 500 times. Several simulations, validation, and characterization experiments were performed on the developed pump to establish its functionality and suitability for use in POCD applications.
Microfluidic In-Flow Decantation Technique Using Stepped Pillar Arrays and Hydraulic Resistance Tuners
Separating the particles from the liquid component of sample solutions is important for several microfluidic-based sample preparations and/or sample handling techniques, such as plasma separation from whole blood, sheath-free flow focusing, particle enrichment etc. This paper presents a microfluidic in-flow decantation technique that provides the separation of particles from particle-free fluid while in-flow. The design involves the expansion of sample fluid channel in lateral and depth directions, thereby producing a particle-free layer towards the walls of the channel, followed by gradual extraction of this particle-free fluid through a series of tiny openings located towards one-end of the depth-direction. The latter part of this design is quite crucial in the functionality of this decantation technique and is based on the principle called wee-extraction. The design, theory, and simulations were presented to explain the principle-of-operation. To demonstrate the proof-of-principle, the experimental characterization was performed on beads, platelets, and blood samples at various hematocrits (2.5%–45%). The experiments revealed clog-free separation of particle-free fluid for at least an hour of operation of the device and demonstrated purities close to 100% and yields as high as 14%. The avenues to improve the yield are discussed along with several potential applications.
Design and validation of on-chip planar mixer based on advection and viscoelastic effects
Mixing at low Reynolds number is usually due to diffusion and requires longer channel lengths for complete mixing. In order to reduce the mixing lengths, advective flow can be induced by varying the channel geometry. Additionally, in non-newtonian fluids, appropriate modifications to channel geometry can be used to aid the mixing process by capitalizing on their viscoelastic nature. Here we have exploited the advection and viscoelastic effects to implement a planar passive micro-mixer. Microfluidic devices incorporating different blend of mixing geometries were conceived. The optimum design was chosen based on the results of the numerical simulations performed in COMSOL. The chosen design had sudden expansion and contraction along with teeth patterns along the channel walls to improve mixing. Mixing of two different dyes was performed to validate the mixing efficiency. Particle dispersion experiments were also carried out. The results indicated effective mixing. In addition, the same design was also found to be compatible with electrical power free pumping mechanism like suction. The proposed design was then used to carry out on-chip chemical cell lysis with human whole blood samples to establish its use with non-newtonian fluids. Complete lysis of the erythrocytes was observed leaving behind the white blood cells at the outlet.
In situ monitoring of photostriction in chalcogenide glass film using fiber Bragg grating sensors
The reversible photostriction (photomechanical strain) in Ge 35 S 65 chalcogenide thin film deposited by a solvent casting method has been monitored using a fiber Bragg grating (FBG) sensor. The shift in Bragg wavelength is used as a probing parameter to quantitatively measure the photoinduced strain arising because of structural modifications in these films under illumination. Exposure to band gap light (405 nm) and above band gap light (302 and 254 nm) leads to a reversible photostriction effect of the order of 100 µε. The present study shows that FBG sensors can be used to effectively measure the optomechanical actuation in chalcogenide films caused by the reversible photostriction effect in the visible and ultraviolet wavelength region.
Photocatalytic degradation of MB by TiO2: studies on recycle and reuse of photocatalyst and treated water for seed germination
Photocatalysis is an effective way for treatment of wastewater and degradation of dyes. It is important to assess the reusability of photocatalyst and treated water after the treatment process. In this study, the photocatalytic activity of TiO 2 (titanium dioxide) and TiO 2 -TMAOH (titanium dioxide-tetramethylammonium hydroxide) was analyzed for degradation of methylene blue dye. Enhanced degradation of methylene blue is observed while treated with TiO 2 -TMAOH with photodegradation efficiency (PDE) 80% within 20 min. A further study shows the reusability of TiO 2 for degradation of dye for six cycles with a decrease in photodegradation efficiency from 90% (cycle-1) to 50% (cycle-2). Fourier transform infrared spectroscopy (FTIR), energy-dispersive X-ray spectroscopy (EDX), and cyclic voltammetry (CV) analysis were carried out to identify the functional groups in treated water, traces of titanium, and TMAOH, respectively. Seed germination of Vigna radiata using TiO 2 - and TiO 2 -TMAOH-treated water shows equivalent and consistent growth. Water quality analysis of treated water shows improved biochemical oxygen demand (BOD) level (1.5 mg L −1 ), which is suitable for reusability of water for many applications. The outcomes suggest treated water can be used for irrigation and plantation purposes.
Microwave irradiation-based rapid curing of PDMS for microfluidic applications
Microfluidic devices are routinely fabricated using replica moulding. The master relief structure is generally constructed on a silicon substrate and the stamps are made using polydimethylsiloxane (PDMS), cured via the slow process of hot-air convective heating. An alternative method that greatly reduces this cure time is presented here. It employs a low-cost, consumer-grade microwave oven and speeds up the curing process by permitting a much higher heat flux to be applied to the prepolymer. A specific pulsed irradiation protocol has been devised which achieves precisely controlled energy delivery for this purpose. This is necessary because silicon’s relatively much stronger microwave absorption causes uncontrollable temperature overshoots that could degrade PDMS if continuously irradiated. The protocol has been arrived at after due optimization of operational parameters, achieved via simulation of a finite element model. It has been implemented by modifying the control circuitry of the consumer grade oven. Cure times as low as two minutes have been demonstrated. Optical and mechanical characterization results show no adverse effect on the performance of PDMS cured using the proposed method. Thus, it can be used for the rapid fabrication of PDMS-based microfluidic devices and would be especially beneficial for research labs looking to speed up microfluidic device development iterations.
Rapid diagnosis of Leishmania infection with a portable loop-mediated isothermal amplification device
L. donovani is an intracellular protozoan parasite, that causes visceral leishmaniasis (VL), and consequently, post-kala azar dermal leishmaniasis (PKDL). Diagnosis and treatment of leishmaniasis is crucial for decreasing its transmission. Various diagnostic techniques like microscopy, enzyme-linked immunosorbent assays (ELISA) and PCR-based methods are used to detect leishmaniasis infection. More recently, loop-mediated isothermal amplification (LAMP) assay has emerged as an ideal diagnostic measure for leishmaniasis, primarily due to its accuracy, speed and simplicity. However, point-of-care diagnosis is still not been tested with the LAMP assay. We have developed a portable LAMP device for the monitoring of Leishmania infection. The LAMP assay performed using our device can detect and amplify as little as 100 femtograms of L. donovani DNA. In a preliminary study, we have shown that the device can also amplify L. donovani DNA present in VL and PKDL patient samples with high sensitivity (100%), specificity (98%) and accuracy (99%), and can be used both for diagnostic and prognostic analysis. To our knowledge, this is the first report to describe the development and application of a portable LAMP device which has the potential to evolve as a point-of-care diagnostic and prognostic tool for Leishmania infections in future.
Microfluidic microscopy-assisted label-free approach for cancer screening: automated microfluidic cytology for cancer screening
Each year, about 7–8 million deaths occur due to cancer around the world. More than half of the cancer-related deaths occur in the less-developed parts of the world. Cancer mortality rate can be reduced with early detection and subsequent treatment of the disease. In this paper, we introduce a microfluidic microscopy-based cost-effective and label-free approach for identification of cancerous cells. We outline a diagnostic framework for the same and detail an instrumentation layout. We have employed classical computer vision techniques such as 2D principal component analysis-based cell type representation followed by support vector machine-based classification. Analogous to criminal face recognition systems implemented with help of surveillance cameras, a signature-based approach for cancerous cell identification using microfluidic microscopy surveillance is demonstrated. Such a platform would facilitate affordable mass screening camps in the developing countries and therefore help decrease cancer mortality rate.
Handheld Fluorescence Microscopy based Flow Analyzer
Fluorescence microscopy has the intrinsic advantages of favourable contrast characteristics and high degree of specificity. Consequently, it has been a mainstay in modern biological inquiry and clinical diagnostics. Despite its reliable nature, fluorescence based clinical microscopy and diagnostics is a manual, labour intensive and time consuming procedure. The article outlines a cost-effective, high throughput alternative to conventional fluorescence imaging techniques. With system level integration of custom-designed microfluidics and optics, we demonstrate fluorescence microscopy based imaging flow analyzer. Using this system we have imaged more than 2900 FITC labeled fluorescent beads per minute. This demonstrates high-throughput characteristics of our flow analyzer in comparison to conventional fluorescence microscopy. The issue of motion blur at high flow rates limits the achievable throughput in image based flow analyzers. Here we address the issue by computationally deblurring the images and show that this restores the morphological features otherwise affected by motion blur. By further optimizing concentration of the sample solution and flow speeds, along with imaging multiple channels simultaneously, the system is capable of providing throughput of about 480 beads per second.