Enzymatic degradation of bacterial cellulose derived carbon nanofibers (BC-CNF) by myeloperoxidase (MPO): Performance evaluation for biosensing

Nandeshwar, Ruchira; Illa, Mani Pujitha; Khandelwal, Mudrika; Tallur, Siddharth

doi:10.1016/j.biosx.2022.100252

Cited by 4 publications

(6 citation statements)

References 49 publications

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“…The RNA extracted was used as template for further PCR reactions. Detailed sample preparation procedure is reported in our earlier work [5]. Amplification was carried out parallelly in WELPCR and a commercially available thermocycler (Bio-Rad T100™ Thermal Cycler) to compare the performance.…”

Section: Resultsmentioning

confidence: 99%

“…cDNA was synthesized on the both instruments using the iScript cDNA synthesis kit (Bio-Rad Laboratories) [6] as per the manufacturer’s protocol. Briefly, it consists of three steps: priming at 25 ° C for 5 min, reverse transcription at 46 ° C for 20 min and inactivation of reverse trancriptase enzyme at 95 ° C for 1 min [5]. Synthesised cDNA was used as a template for amplifying a 503 bp long fragment.…”

Section: Resultsmentioning

confidence: 99%

“…For demonstrating the utility of WELPCR, we amplified nucleic acid from bacteriophage phi6, an enveloped doublestranded RNA (dsRNA) virus that infects Pseudomonas syringae (non-pathogenic BSL-1 strain). This virus serves as a surrogate for highly pathogenic enveloped viruses such as SARS-CoV-2, ebola and influenza [4], [5]. Using GenE-lute™ Universal Total RNA purification kit (Sigma-Aldrich), RNA extraction was carried out as per the manufacturer's protocol.…”

Section: B Sample Preparationmentioning

confidence: 99%

“…Using GenE-lute™ Universal Total RNA purification kit (Sigma-Aldrich), RNA extraction was carried out as per the manufacturer's protocol. Extracted RNA was quantified by determining the absorbance at 260 nm using a UV/Vis spectrophotometer (Thermo Scientific Multiskan GO) [5]. The RNA extracted was used as template for further PCR reactions.…”

Section: B Sample Preparationmentioning

confidence: 99%

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WELPCR: Low-cost polymerase chain reaction (PCR) thermal cycler for nucleic acid amplification and sensing

Mandal,

Dhakane,

Kulkarni

et al. 2023

Preprint

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The COVID-19 pandemic highlighted the need for sensitive and cost-effective diagnostics. The gold standard test to diagnose infectious diseases such as COVID-19 is quantitative polymerase chain reaction (qPCR). Although there have been multiple advances in nucleic acid testing, PCR thermocyclers are expensive and typically not affordable for offering hands-on training to PCR in instructional laboratory courses in undergraduate colleges. We introduce a low-cost, portable, and automated real-time PCR device designed at the Wadhwani Electronics Lab (WEL) at IIT Bombay, christened WELPCR. WELPCR is an open-source design, and capable of reverse transcription PCR (RT-PCR) for complementary DNA synthesis, as well as end-point PCR of small amount of DNA in a test sample. The WELPCR design enables ramp rate up to 2 °C/s, weighs less than 2 kg, and is easy to assemble. The built-in LED display and keypad provides an intuitive and seamless user interface to configure and set up WELPCR. With bill of materials (BOM) cost less than INR Rs. 10 000, WELPCR is an attractive solution for bringing PCR technology to instructional and research laboratories in a cost-effective manner.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: B Sample Preparationmentioning

confidence: 99%

Section: B Sample Preparationmentioning

confidence: 99%

See 2 more Smart Citations

WELPCR: Low-cost polymerase chain reaction (PCR) thermal cycler for nucleic acid amplification and sensing

Mandal,

Dhakane,

Kulkarni

et al. 2023

Preprint

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show abstract

“…In this respect, in the last few years the enzymatic degradation of carbon nanostructures has been extensively analyzed in the literature. [1][2][3][4][5] This is mainly due to a large number of publications speculating about the possible uses of fullerenes, nanotubes, and graphene in biomedical applications, for example, as delivering therapeutic agents, 6 for the development of new diagnostic strategies, 7 tissue engineering, 8,9 biosensors, 10,11 and so on. Consequently, not only toxicity studies 12,13 but also efforts to develop strategies to achieve their complete elimination from the human body become crucial.…”

Section: Introductionmentioning

confidence: 99%

ClO-driven degradation of graphene oxide: new insights from DFT calculations

Romo-Ávila,

Márquez-Ruíz,

Guirado-López

2024

Phys. Chem. Chem. Phys.

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Electrochemical detection of myeloperoxidase (MPO) in blood plasma with surface-modified electroless nickel immersion gold (ENIG) printed circuit board (PCB) electrodes

Nandeshwar,

Tallur

2023

Preprint

Self Cite

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Printed circuit board (PCB) based biosensors have often utilized hard gold electroplating, that nullifies the cost advantages of this technology as compared to screen printed electrodes. Electroless nickel immersion gold (ENIG) is a popular gold deposition process widely used in PCB manufacturing, but vulnerable to pinhole defects and large surface roughness, which compromises biosensor performance. In this work, we present a method to address these challenges through electrodeposition of methylene blue (MB) to cover surface defects and improve electroactivity of ENIG PCB electrodes. We also demonstrate a process to realize in situ synthesis of gold nanoparticles (AuNPs) using acid-functionalized multi-walled carbon nanotubes (MWCNTs) as scaffold, that are used to immobilize antibody for the target molecule (myeloperoxidase: MPO, early warning biomarker for cardiovascular diseases) through a modified cysteamine/gluteraldehyde based process. The processing steps on the electrode surface are developed in a manner that do not compromise the integrity of the electrode, resulting in repeatable and reliable performance of the sensors. Further, we demonstrate a cost-effective microfluidic packaging process to integrate a capillary pump driven microfluidic channel on the PCB electrode for seamless introduction of samples for testing. We demonstrate the ability of the sensor to distinguish clinically abnormal concentrations of MPO from normal concentrations through extensive characterization using spiked serum and blood plasma samples, with a limit of detection of 0.202 ng/mL.

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Enzymatic degradation of bacterial cellulose derived carbon nanofibers (BC-CNF) by myeloperoxidase (MPO): Performance evaluation for biosensing

Cited by 4 publications

References 49 publications

WELPCR: Low-cost polymerase chain reaction (PCR) thermal cycler for nucleic acid amplification and sensing

WELPCR: Low-cost polymerase chain reaction (PCR) thermal cycler for nucleic acid amplification and sensing

ClO-driven degradation of graphene oxide: new insights from DFT calculations

Electrochemical detection of myeloperoxidase (MPO) in blood plasma with surface-modified electroless nickel immersion gold (ENIG) printed circuit board (PCB) electrodes

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