Point of care real-time polymerase chain reaction-based diagnostic for Kyasanur forest disease

Majumdar, Triparna; Shete, Anita M.; Yadav, Pragya D; Patil, Savita; Mali, Deepak; Waghmare, Ashwini; Gawande, Pranita

doi:10.1016/j.ijid.2021.05.036

Cited by 5 publications

(3 citation statements)

References 12 publications

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“…[40] For comparison of the proposed biosensor to already established techniques, data is taken from published research data . [35][36][37][38][39] Archive Clinical data in which the viral load of CHIKV is also published is taken to compare the results of the proposed biosensor. One hundred and seven archive results of the RT-PCR test were used to determine accuracy measures of MEMS Biosensor.…”

Section: Comparison With the Clinical Datamentioning

confidence: 99%

Polymer‐Based MEMS Sensor for Label‐Free Chikungunya Virus Detection

Niranjan

Gupta

Rajoria

2023

Macromolecular Symposia

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Piezoelectric MEMS (Micro-Electro-Mechanical Systems) Cantilevers have successfully been used in a wide variety of applications in recent years. This paper is a simulation-based study on Micro-cantilever based Piezoelectric MEMS biosensors for Chikungunya Virus (CHIKV) detection. In addition to providing early detection, the proposed method is label-free, faster, and more reliable. The proposed biosensor detects CHIKV E2 protein via MEMS Sensor. The basic principle involves using piezoelectric material on a micro-cantilever for biomaterial detection. A piezoelectric MEMS cantilever is simulated in COMSOL Multiphysics and compared for voltage, stress, and displacement for three materials (silicon, gold, and PDMS). Analytical equations are derived. Post-processing for biosensor design is done in SIMULINK. Simulated voltage, stress, and displacement show a linear relationship with the viral load. PDMS also provided the highest voltage and displacement among the three materials used for MEMS simulation. The graphs show the comparison between simulated and analytical values. Simulation results are also compared to clinical results to prove the biosensor's validity. MEMS Biosensor performance matches with the RT-PCR results, with the added advantage of faster results. Diagnostic tests in the medical field can be performed using the proposed biosensor.

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Section: Comparison With the Clinical Datamentioning

confidence: 99%

Polymer‐Based MEMS Sensor for Label‐Free Chikungunya Virus Detection

Niranjan

Gupta

Rajoria

2023

Macromolecular Symposia

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“…In severe cases, neurological and haemorrhagic manifestations are observed (7), with a mortality of ∼ 2-10% (5,7). ICMR-NIV and Molbio Diagnostics, Goa, have developed a microchip-based point-of-care test (Truenat KFD), which is currently the gold standard for diagnosis in India (8,9). Formalin inactivated tissue-culture vaccine is available for KFD, which is administered in two doses to individuals aged 7–65 years, at one month interval (10).…”

Section: Introductionmentioning

confidence: 99%

“…In extreme cases, neurological and hemorrhagic manifestations are observed, with an estimated fatality of ~ 3-5% [6]. A microchip-based point-of-care test (Truenat KFD), developed by ICMR-NIV and Molbio Diagnostics, Goa, is currently the gold standard for diagnosis in India [7]. Formalin inactivated tissue-culture vaccine is available for KFD, administered in two doses to individuals aged 7–65 years at one-month intervals [8].…”

Section: Introductionmentioning

confidence: 99%

Sofosbuvir and its tri-phosphate metabolite inhibit the RNA-dependent RNA polymerase activity of non-Structural protein 5 from the Kyasanur forest disease virus

Malik

Vijayan

Jagannath

et al. 2022

Preprint

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Kyasanur forest disease is a neglected zoonotic disease caused by a single stranded RNA based flavivirus, the incidence of which, was first recorded in 1957, in the Southern part of India. Kyasanur forest disease virus is transmitted to monkeys and humans, through the infected tick bite of Haemophysalis spinigera. Kyasanur forest disease is a febrile illness, which in severe cases, results in neurological manifestations leading to mortality. The current treatment regimens are symptomatic and supportive in nature. There are no targeted therapies available for this disease. In this study, we evaluated the ability of the FDA approved drug, Sofosbuvir to inhibit the RNA dependent RNA polymerase activity of NS5 protein from Kyasanur forest disease virus. NS5 protein containing the N-terminal methyl transferase domain and C-terminal RNA dependent RNA polymerase domain was expressed in Escherichia coli and RNA dependent RNA polymerase activity, demonstrated with the purified protein. The RNA dependent RNA polymerase assay conditions were optimized, followed by determination of apparent Km,ATP. IC50 of Sofosbuvir against RNA dependent RNA polymerase activity was determined under the optimized conditions with the purified NS5 protein. Drug repurposing strategies can accelerate the development of targeted therapies for Kyasanur forest disease. This is the first demonstration of inhibition of RNA dependent RNA polymerase activity of Kyasanur forest disease virus with a small molecule inhibitor.

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