Development of a Quantitative Real-Time PCR Assay for Detection of Toxoplasma gondii in Brain Samples

Background: Toxoplasmosis is a worldwide-distributed infection that can cause serious diseases, mainly in congenitally infected and immunodeficient individuals. PCR assays play an indispensable role in the detection of Toxoplasma gondii in different biological samples. Methods: This study was conducted in the Parasitology Department at Pasteur Institute of Iran (Tehran) during 2016-2018. We designed a highly sensitive quantitative real-time PCR (RT-qPCR) targeted REP-529, a noncoding repetitive DNA. We cloned the amplicon in a plasmid (pTZREP-529) and used it to generate the standard curve. The Toxoplasma RT-qPCR characteristics, i.e., detection limit, specificity, linear dynamic range, linearity, intra-, and inter-assay precisions, were determined. The detection limit of the assay was one plasmid copy number (PCN) per reaction (about 0.004 T. gondii genome), and the linear dynamic range was equal to 6 logs (1× 101 to 1× 107 PCN per reaction). Results: The assay showed no signal when genomic DNA of Plasmodium falciparum, Leishmania major, and Trichomonas vaginallis were used. The standard curve was drawn using dilutions of pTZREP-529 plasmid spiked with genomic DNA from a mouse brain, and test characteristics were shown unaffected. Applying the Toxoplasma RT-qPCR, we showed brain cysts were significantly decreased in mice vaccinated with GRA2 antigen of Toxoplasma formulated in Monophosphoryl Lipid A (MPL) adjuvant. Conclusion: We have developed a quantitative, specific, and highly sensitive PCR for detecting T. gondii in biological samples.

Screen-Printed Electrode Surface Modification with NiCo2O4/RGO Nanocomposite for Hydroxylamine Detection

We developed a novel hydroxylamine sensor through the surface modification of screen-printed electrode (SPE) with NiCo2O4 nanoparticles/reduced graphene oxide (RGO) nanocomposite (NiCo2O4/RGO/SPE). We assessed the electrochemical response of hydroxylamine on the as-fabricated sensor, confirming the high electrocatalytic impact of hydroxylamine oxidation. The electrode produced sensitively responded to hydroxylamine under optimized conditions, with a low limit of detection (2.0 nM) and broad linear dynamic range (0.007–385.0 µM). The presence of NiCo2O4 combined with the modification of RGO resulted in sensitive detection and signal amplification of hydroxylamine oxidation. The proposed sensor was used to determine the existence of hydroxylamine in water samples.

Quantum-assisted distortion-free audio signal sensing

Quantum sensors are keeping the cutting-edge sensitivities in metrology. However, for high-sensitive measurements of arbitrary signals, limitations in linear dynamic range could introduce distortions when sensing the frequency, magnitude and phase of unknown signals. Here, we overcome these limitations with advanced sensing protocol that combines quantum phase-sensitive detection with the heterodyne readout. We present theoretical and experimental investigations using nitrogen-vacancy centers in diamond, showing the ability to sense audio signals with a 98 dB linear dynamic range, a 31 pT/Hz1/2 sensitivity, and arbitrary frequency resolution. Further, we perform the quantum-assisted distortion-free audio signal (melody, speech) sensing with high fidelity. The methods developed here could broaden the horizon for quantum sensors towards applications in telecommunication, where high-fidelity and low-distortion at multiple frequency bands within small sensing volumes are required.

Novel hydrophilic-phase extraction, HILIC and high-resolution MS quantification of an RNA oligonucleotide in plasma

Aim: In the theme of quantitative LC–MS bioanalysis of oligonucleotides free of ion-pairing, a 22-mer RNA oligonucleotide took center stage. The focus was on a unique polar-based retention scheme to produce a high-recovery extraction presenting a high-performance alternative extraction means, also there was the opportunity to involve hydrophilic-interaction liquid chromatography and contemporary high-resolution MS as the end point. Results: Original LC–MS methodology was developed for the oligonucleotide and the performance was robust for both nominal and accurate mass detection, the latter affording 10× improvement in sensitivity and 4000-fold linear dynamic range, 500 pM to 2000 nM. Conclusion: A novel means of solid-phase extraction is exhibited within a robust pair-free methodology, reaching pM sensitivity with the demonstrably beneficial accurate mass platform.

Photodetector Based on CsPbBr3/Cs4PbBr6 Composite Nanocrystals with High Detectivity

High-quality, all-inorganic CsPbBr3/Cs4PbBr6 composite perovskite nanocrystals (NCs) were obtained with all-solution-processing at room temperature, and a photodetector (PD) with high detectivity was realized based on CsPbBr3/Cs4PbBr6 NCs. The detectivity (D*) of the proposed PD is 4.24 × 1012 Jones under 532 nm illumination, which is among the highest levels for PDs based on all-inorganic perovskite NCs. In addition, a high linear dynamic range (LDR) of 115 dB under 1 V bias was also realized. Furthermore, the underlying mechanism for the enhanced performance of the proposed PD was discussed. Our work might promote the preparation of high-performance PDs based on dual-phase all-inorganic perovskite nanocrystals.