scholarly journals Evaluation of a Lyophilized CRISPR-Cas12 Assay for a Sensitive, Specific, and Rapid Detection of SARS-CoV-2

Viruses ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 420
Author(s):  
Lucía Ana Curti ◽  
Ivana Primost ◽  
Sofia Valla ◽  
Daiana Ibañez Alegre ◽  
Cecilia Olguin Perglione ◽  
...  
Keyword(s):  
Respiratory Pathogens ◽  
Kappa Index ◽  
Test Procedures ◽  
Validation Test ◽  
Initial Validation ◽  
Highly Sensitive ◽  
Wide Range ◽  
Sophisticated Equipment ◽  
Lysis Buffer ◽  
Threshold Range

We evaluated a lyophilized CRISPR-Cas12 assay for SARS-CoV-2 detection (Lyo-CRISPR SARS-CoV-2 kit) based on reverse transcription, isothermal amplification, and CRISPR-Cas12 reaction. From a total of 210 RNA samples extracted from nasopharyngeal swabs using spin columns, the Lyo-CRISPR SARS-CoV-2 kit detected 105/105 (100%; 95% confidence interval (CI): 96.55–100) positive samples and 104/105 (99.05%; 95% CI: 94.81–99.97) negative samples that were previously tested using commercial RT-qPCR. The estimated overall Kappa index was 0.991, reflecting an almost perfect concordance level between the two diagnostic tests. An initial validation test was also performed on 30 nasopharyngeal samples collected in lysis buffer, in which the Lyo-CRISPR SARS-CoV-2 kit detected 20/21 (95.24%; 95% CI: 76.18–99.88) positive samples and 9/9 (100%; 95% CI: 66.37–100) negative samples. The estimated Kappa index was 0.923, indicating a strong concordance between the test procedures. The Lyo-CRISPR SARS-CoV-2 kit was suitable for detecting a wide range of RT-qPCR-positive samples (cycle threshold range: 11.45–36.90) and dilutions of heat-inactivated virus (range: 2.5–100 copies/µL); no cross-reaction was observed with the other respiratory pathogens tested. We demonstrated that the performance of the Lyo-CRISPR SARS-CoV-2 kit was similar to that of commercial RT-qPCR, as the former was highly sensitive and specific, timesaving (1.5 h), inexpensive, and did not require sophisticated equipment. The use of this kit would reduce the time taken for diagnosis and facilitate molecular diagnosis in low-resource laboratories.

Preparation of Functionalized Diorganomagnesium Reagents in Toluene via Bromine or Iodine/Magnesium-Exchange Reactions

Synthesis ◽  
2021 ◽  
Author(s):  
Alexandre Desaintjean ◽  
Fanny Danton ◽  
Paul Knochel
Keyword(s):  
Exchange Reaction ◽  
Nitro Group ◽  
General Formula ◽  
Functional Groups ◽  
Exchange Reactions ◽  
Acyl Chlorides ◽  
Aryl Iodides ◽  
Highly Sensitive ◽  
Wide Range ◽  
Magnesium Exchange

A wide range of polyfunctionalized di(hetero)aryl- and dialkenyl-magnesium reagents were prepared in toluene within 10 to 120 min between −78 °C and 25 °C via an I/Mg- or Br/Mg-exchange reaction using reagents of the general formula R2Mg (R = sBu, Mes). Highly sensitive functional groups, such as a triazene or a nitro group, were tolerated in these exchange reactions, enabling the synthesis of various functionalized (hetero)arenes and alkenes derivatives after quenching with several electrophiles including allyl bromides, acyl chlorides, aldehydes, ketones, and aryl iodides.

scholarly journals 6-Axis Stress Tensor Sensor Using Multifaceted Silicon Piezoresistors

Micromachines ◽  
2021 ◽  
Vol 12 (3) ◽  
pp. 279
Author(s):  
Kentaro Noda ◽  
Jian Sun ◽  
Isao Shimoyama
Keyword(s):  
Stress Tensor ◽  
Elastic Body ◽  
Building Materials ◽  
Sensor Response ◽  
Stress Change ◽  
Sensor Chip ◽  
Naked Eye ◽  
Highly Sensitive ◽  
Wide Range

A tensor sensor can be used to measure deformations in an object that are not visible to the naked eye by detecting the stress change inside the object. Such sensors have a wide range of application. For example, a tensor sensor can be used to predict fatigue in building materials by detecting the stress change inside the materials, thereby preventing accidents. In this case, a sensor of small size that can measure all nine components of the tensor is required. In this study, a tensor sensor consisting of highly sensitive piezoresistive beams and a cantilever to measure all of the tensor components was developed using MEMS processes. The designed sensor had dimensions of 2.0 mm by 2.0 mm by 0.3 mm (length by width by thickness). The sensor chip was embedded in a 15 mm3 cubic polydimethylsiloxane (PDMS) (polydimethylsiloxane) elastic body and then calibrated to verify the sensor response to the stress tensor. We demonstrated that 6-axis normal and shear Cauchy stresses with 5 kPa in magnitudes can be measured by using the fabricated sensor.

scholarly journals State of the Science in Dried Blood Spots

2018 ◽  
Vol 64 (4) ◽  
pp. 656-679 ◽  
Author(s):  
Jeffrey D Freeman ◽  
Lori M Rosman ◽  
Jeremy D Ratcliff ◽  
Paul T Strickland ◽  
David R Graham ◽  
...  
Keyword(s):  
Systematic Approach ◽  
Full Range ◽  
Dried Blood Spots ◽  
Blood Spots ◽  
Highly Sensitive ◽  
Wide Range ◽  
Potential Applications ◽  
Review Of Reviews ◽  
Near Term ◽  
State Of The Science

Abstract BACKGROUND Advancements in the quality and availability of highly sensitive analytical instrumentation and methodologies have led to increased interest in the use of microsamples. Among microsamples, dried blood spots (DBS) are the most well-known. Although there have been a variety of review papers published on DBS, there has been no attempt at describing the full range of analytes measurable in DBS, or any systematic approach published for characterizing the strengths and weaknesses associated with adoption of DBS analyses. CONTENT A scoping review of reviews methodology was used for characterizing the state of the science in DBS. We identified 2018 analytes measured in DBS and found every common analytic method applied to traditional liquid samples had been applied to DBS samples. Analytes covered a broad range of biomarkers that included genes, transcripts, proteins, and metabolites. Strengths of DBS enable its application in most clinical and laboratory settings, and the removal of phlebotomy and the need for refrigeration have expanded biosampling to hard-to-reach and vulnerable populations. Weaknesses may limit adoption in the near term because DBS is a nontraditional sample often requiring conversion of measurements to plasma or serum values. Opportunities presented by novel methodologies may obviate many of the current limitations, but threats around the ethical use of residual samples must be considered by potential adopters. SUMMARY DBS provide a wide range of potential applications that extend beyond the reach of traditional samples. Current limitations are serious but not intractable. Technological advancements will likely continue to minimize constraints around DBS adoption.

scholarly journals Multi-Quantum Dots-Embedded Silica-Encapsulated Nanoparticle-Based Lateral Flow Assay for Highly Sensitive Exosome Detection

Nanomaterials ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 768
Author(s):  
Hyung-Mo Kim ◽  
Chiwoo Oh ◽  
Jaehyun An ◽  
Seungki Baek ◽  
Sungje Bock ◽  
...  
Keyword(s):  
Quantum Dots ◽  
Limit Of Detection ◽  
Specific Binding ◽  
Calf Serum ◽  
Lateral Flow ◽  
Lateral Flow Immunoassay ◽  
Uniform Size ◽  
Highly Sensitive ◽  
Wide Range ◽  
New Biomarkers

Exosomes are attracting attention as new biomarkers for monitoring the diagnosis and prognosis of certain diseases. Colorimetric-based lateral-flow assays have been previously used to detect exosomes, but these have the disadvantage of a high limit of detection. Here, we introduce a new technique to improve exosome detection. In our approach, highly bright multi-quantum dots embedded in silica-encapsulated nanoparticles (M–QD–SNs), which have uniform size and are brighter than single quantum dots, were applied to the lateral flow immunoassay method to sensitively detect exosomes. Anti-CD63 antibodies were introduced on the surface of the M–QD–SNs, and a lateral flow immunoassay with the M–QD–SNs was conducted to detect human foreskin fibroblast (HFF) exosomes. Exosome samples included a wide range of concentrations from 100 to 1000 exosomes/µL, and the detection limit of our newly designed system was 117.94 exosome/μL, which was 11 times lower than the previously reported limits. Additionally, exosomes were selectively detected relative to the negative controls, liposomes, and newborn calf serum, confirming that this method prevented non-specific binding. Thus, our study demonstrates that highly sensitive and quantitative exosome detection can be conducted quickly and accurately by using lateral immunochromatographic analysis with M–QD–SNs.

Highly Sensitive and Wide Range Non-Enzymatic Electrochemical Detection of Cholesterol using Pencil Lead Electrodes

2021 ◽  
Author(s):  
Navaneeth Punnakkal ◽  
Jeethu Raveendran ◽  
Suneesh Punathil Vasu ◽  
Bipin G Nair ◽  
T G Satheesh Babu
Keyword(s):  
Electrochemical Detection ◽  
Highly Sensitive ◽  
Wide Range ◽  
Lead Electrodes ◽  
Pencil Lead

Broadening the valid temperature range of optical thermometry through dual-mode design

2018 ◽  
Vol 6 (41) ◽  
pp. 11178-11183 ◽  
Author(s):  
Yan Gao ◽  
Yao Cheng ◽  
Tao Hu ◽  
Zeliang Ji ◽  
Hang Lin ◽  
...  
Keyword(s):  
Dual Mode ◽  
Temperature Range ◽  
Mode Design ◽  
Optical Thermometry ◽  
Highly Sensitive ◽  
Wide Range ◽  
Optical Thermometer

This study highlights a highly sensitive dual-mode optical thermometer Pr3+:Gd2ZnTiO6 for thermal readings over a wide range of temperature.

A Methodology for Assessing Fuel Cell Performance Under a Wide Range of Operational Conditions: Results for Single Cells

2006 ◽  
Vol 3 (3) ◽  
pp. 226-233 ◽  
Author(s):  
Andrea Baratella ◽  
Roberto Bove ◽  
Piero Lunghi
Keyword(s):  
Fuel Cells ◽  
Fuel Cell ◽  
Single Cells ◽  
Cell Performance ◽  
Effective Parameters ◽  
Test Procedures ◽  
Operational Conditions ◽  
Fuel Cell Performance ◽  
Wide Range ◽  
The University

Testing the performance of fuel cells is an important key for verifying technology improvements and for demonstrating their potential. However, due to the novelty of this technology, there is not a standardized procedure for testing fuel cell performance. In order to fully investigate fuel cell performance, the behavior must be known under a wide range of operational conditions. Furthermore, in order to compare results coming from different test teams, a set of procedures and parameters to evaluate single cell performance should be defined. The research group of the Fuel Cell Laboratory of the University of Perugia is conducting performance tests on single cells, focusing on defining test procedures to find effective parameters to be used to compare tests performed by different teams. This work demonstrates how the testing parameters developed by the team allow one to perform advanced control on test procedures, to understand test results, and to compare them with tests carried out under different operational conditions. The entire analysis is easily conducted by using a single parameter variation hyperspace approach. The experimental results obtained on single fuel cells are reported.

scholarly journals Highly sensitive and multiplexed in situ protein profiling with cleavable fluorescent streptavidin

2019 ◽  
Author(s):  
Renjie Liao ◽  
Diego Mastroeni ◽  
Paul D. Coleman ◽  
Jia Guo
Keyword(s):  
Signal Amplification ◽  
Individual Cell ◽  
Protein Detection ◽  
Protein Profiling ◽  
Detection Sensitivity ◽  
Layer By Layer ◽  
Protein Targets ◽  
Highly Sensitive ◽  
Wide Range

AbstractThe ability to perform highly sensitive and multiplexed in situ protein analysis is crucial to advance our understanding of normal physiology and disease pathogenesis. To achieve this goal, here we develop an approach using cleavable biotin conjugated antibodies and cleavable fluorescent streptavidin (CFS). In this approach, protein targets are first recognized by the cleavable biotin labeled antibodies. Subsequently, CFS is applied to stain the protein targets. Though layer-by-layer signal amplification using cleavable biotin conjugated orthogonal antibodies and CSF, the protein detection sensitivity can be enhanced by at least 10 fold, compared with the existing methods. After imaging, the fluorophores and the biotins unbound to streptavidin are removed by chemical cleavage. The leftover streptavidin is blocked by biotin. Upon reiterative analysis cycles, a large number of different proteins with a wide range of expression levels can be unambiguously detected in individual cell in situ.

Sign in / Sign up

Export Citation Format

Share Document