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The new PMC design is here! Learn more about navigating our updated article layout. The PMC legacy view will also be available for a limited time. Federal government websites often end in. The site is secure. The spread of SARS-CoV-2 virus in the ongoing global pandemic has led to infections of millions of people and losses of many lives. Diagnosis of patients in the early stage infection are so far limited to viral nucleic acid or antigen detection in human nasopharyngeal swab or saliva samples.

Here we developed a method for rapid and direct optical measurement of SARS-CoV-2 virus particles in one step nearly without any sample preparation using a spike protein specific nanoplasmonic resonance sensor. Measurements shown on both generic microplate reader and a handheld smartphone connected device suggest that our low-cost and rapid detection method may be adopted quickly under both regular clinical environment and resource-limited settings.

It is evident that the infection may be effectively reduced and eventually controlled with swift and large-scale testing for early diagnostics Chen et al. The PCR based nucleic acid testing is very sensitive and accurate in early infection diagnostics but it generally requires multiple and lengthy processes including virus lysis, RNA extraction, reverse transcription and amplification and is prone to sample contamination Ai et al. There were some reported and commercially available viral antigen spike S or nucleocapsid N protein detection assays however they all suffered with very poor accuracy, reliability and sensitivity.

Therefore it is a global consensus there is an urgent and tremendous demand for low-cost rapid and reliable SARS-CoV-2 antigen and virus detection which helps to enable timely and affordable point-of-care COVID diagnostics for a very large population. Chang et al. However conventional SPR testing equipment are bulky and not affordable to most research and clinical institutions especially in developing countries and resources limited settings Moran et al.

Therefore the SPR virus detection, although often showed in research labs, rarely becomes a viable method accessible to clinical and point-of-care applications. Here we report a low-cost nanoplasmonic sensor allowing for one-step rapid detection and quantification of the SARS-CoV-2 pseudovirus Fig. The low-cost plasmonic nanocup array sensor chips were made by the method previously reported Dang et al. Owing to the specially designed periodic nanostructures, without any external coupling optics, the plasmon resonance wavelength and intensity change on the virus-capturing sensor surface can be simply observed by transmission light spectroscopy or imaging Soler et al.

Therefore the nanoplasmonic sensor chips can be integrated with microwell plate or microfluidic cuvette, and the measurements are carried out in both ubiquitous generic microplate readers and a low-cost handheld point-of-care testing device Dang et al. Similar sensing capability are demonstrated by using a low-cost handheld optical equipment controlled by a smartphone App. The ultrasensitive SARS-CoV-2 virus detection and potential early diagnosis of COVID disease become available for point-of-care applications in clinics, roadside triage sites and even home settings.

Scanning electron microscopy image Left shows the replicated nanocup array. Transmission microscopy image Right shows that air and water on the device surface exhibit different colors, green and far red pink, respectively.

All chemicals were used as obtained without any further purification. The nanoplasmonic sensor was fabricated by a replica molding process. An optical adhesive liquid was evenly spread on the mold and placed on a polyethylene terephthalate PET sheet. Then, 10 nm of titanium Ti and 70 nm of gold Au were deposited onto the nanocup array in an electron beam evaporator.

After rinsing the chip integrated microplate wells or cards twice with deionized DI water, the chips were incubated in a 10 mM MUA ethanol solution for 0. This was done in as much as the optimum stability of the conjugates is at pH 7. The Au-NPs suspension was then centrifuged at rpm for 30 min.

Centrifugation and resuspension steps were repeated 3 times. The dynamic absorption spectrum corves were measured using a simple small volume microplate reader Xlement SPR, Liangzhun Industrial Co. Ltd, Shanghai, China. The purpose of this study is to develop a nanoplasmonic sensor chip detecting SARS-CoV-2 virus particles and controlling the severe pandemic.

After the replica molding process as described in the experiment section, the periodic nanocup array sensor chip was fabricated on a polymer substrate with nm nanocup diameter, nm depth and nm periodicity and 70 nm-in-thickness Au layer on it.

A high uniformity cup array structure can be seen in the scanning electron microscope SEM image of the sensor chip and each nanocup has the similar morphology Fig.

Dang et al. EOT phenomenon is attributed to the strong field-enhanced plasma excitation in the vicinity of the nanoarrays induced by grating coupling of light at normal incidence illumination, which can simplify optical systems design and generate a high sensitivity to minute local refractive index changes Ebbesen et al.

Based on the advantage of EOT, a low-cost ultrasensitive biosensor was integrated into the standard well plate and chip cartridge for direct monitoring the dynamic binding curves of SARS-CoV-2 pseudovirus and SARS-CoV-2 mAbs only using a generic microplate reader and a low-cost handheld optical equipment controlled by a smartphone App Figure S1 , Supporting Information.

In addition, previous results indicated that gold nanoparticles Au-NPs could significantly enhance the detection sensitivity of the nanoplasmonic biosensor Belushkin et al.

The coupling between nanoplasmonic substrates and AuNPs enables quantification of low concentration analyte in a solution with limited diffusion condition.

Importantly, AuNP-enhanced technique can further amplify the optical signal and shorten the detection time. Meanwhile, the luciferase activity study showed that the relative luciferase units RLU.

Fast and sensitive detection of intact coronavirus directly in body fluid samples are essential to diagnose these viruses and help proper treatment. For this reason, our chip-in-microwell sensor were developed for the direct multichannel detection of different concentrations of whole virus particles rather than genetic extraction and analysis methods, which are accurate, but also time-consuming and labor-intensive Fig.

The surface of the nanoplasmonic array sensor chip was functionalized with SARS-CoVspecific antibodies to capture intact coronavirus by binding with spike proteins on its surface Fig.

The original absorption spectra and differential spectra of adjacent sensor modification steps exhibited obvious change Fig. From the absorption spectra of the nanocup array chip, the specific resonant wavelength could be seen at nm. In the following experiments, we choose nm to investigate the dynamic binding curves and standard curves with respect to the different concentration of SARS-CoV-2 pseudovirus.

The real-time dynamic binding curves of the antigen-antibody interaction was shown in Fig. It can be seen that the curve of the control PBS solution group shows almost no change. The relative OD change value was obtained from the dynamic measurement over 60 min process as shown in Fig. The results were analyzed and fitted using a four-parameter logistic 4 PL equation, and the correlation coefficient R 2 is 0.

Moreover, the R 2 is 1. In addition, we present a gold nanoparticles AuNP -enhanced detection techniques based on our nanoplasmonic resonance sensor device that enables enhanced-sensitivity, rapid and convenience coronavirus detection Fig. The real-time dynamic binding curve of the interaction is represented in Fig. Therefore, we used paired antibody sandwich detection, that is, one kind of S protein antibodies conjugated on the nanocup sensor surface and another kind of S protein antibodies labeled on the AuNP surface to respectively capture two distinctive epitopes of the S proteins on the viral particle surface.

As expected, the single-step double-antibody sandwich plasmonic resonance immunoassay method can not only expand the detection range 0—1. In addition, the results were fit to the 4 PL curve and showed good agreement over the range of 0—1. Similarly, the R 2 also reached 0. Reliable detection of SARS-CoV-2 virus requires distinguishing nonspecific binding of other viruses to the functionalized nanoplasmonic sensor surface. A significant difference in binding capacity was observed with a high response to the SARS-CoV-2 viruses while almost no response to other virus strains Fig.

Demand for rapid, accurate and convenient SARS-CoV-2 virus detection present significant challenges in controlling and stopping the pandemics. Diagnosis of patients in the early stage infection are so far limited to viral nucleic acid or antigen detection in human nasopharyngeal swabs or saliva samples.

Although traditional approaches, including point-of-care POC diagnostics, bedside testing, and community-based approaches, were applied to address these challenges, innovative techniques combining with mobile technologies, nanotechnology, imaging systems, and microfluidic technologies are expected to promote this transformation Im et al.

In this work, we also developed a portable and innovative devices controlled by a smartphone App for real-time measurements of the dynamic binding curves of SARS-CoV-2 virus on the nanoplasmonic sensor Fig.

We integrated the nanoplasmonic sensor chip in a cartridge designed for the handheld testing device, followed by functional modification of the sensor chip and detection of pseudovirus particle samples according to the protocol described previously Fig.

The functionalized chip cartridge with different concentrations of pseudovirus samples was inserted into the testing device and the dynamic curves were recorded in real time through the smartphone APP. The real-time virus binding curve measurement is presented in Fig.

This low-cost handheld sensing platform can directly detect the SARS-CoV-2 pseudovirus sample in one step within 15 min and the detectable virus concentrations range over 0 to 6.

Moreover, the quantification limit of the handheld equipment is currently about SARS-CoV-2 virus particles and can be further improved to be comparable with the microplate reader case. As shown in Fig. A remarkably rising curve was observed only for the SARS-CoV-2 pseudovirus sample, suggesting that the devices controlled by a smartphone App could offer convenient operability while permit highly sensitive and specific detection of SARS-CoV-2 pseudovirus.

This nanoplasmonic sensor device with the potential in rapid and affordable early diagnosis of COVID disease will become available for POC applications in clinics, roadside triage site and even home settings. However, there are just a limited rapid diagnostic methods or testing equipments that are effective for newly infected patients or asymptomatic carriers. Therefore in this work, we developed a simple and low-cost device to rapidly and sensitively detect the SARS-CoV-2 virus in one step using a nanoplasmonic biosensor integrated with a standard well plate or a chip cartridge.

Moreover, similar rapid and sensitive detection capabilities are demonstrated by using a low-cost handheld optical equipment controlled by a smartphone App. The quantification limit of SARS-CoV-2 pseudovirus in the handheld system was about virus particles within 15 min.

Longfei Ding: Pseudovirus production and quantification. Jun Zhou: Chip manufacturing, Pseudovirus design and antibody selection. Shuiliang Chen: Chip manufacturing. Fang Chen: Chip manufacturing. Chen Zhao: Investigation, Resources. Jianqing Xu: Medical advisory. Wenjun Hu: Supervision, Funding acquisition.

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. The following are the supplementary data related to this article:. Biosens Bioelectron. Published online Oct Gang L.

Author information Article notes Copyright and License information Disclaimer. Ltd, Wuhan, China. Liangzhun Shanghai Industrial Co. Zhejiang University, Zhejiang, China. All rights reserved. Elsevier hereby grants permission to make all its COVIDrelated research that is available on the COVID resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source.

This article has been cited by other articles in PMC. Associated Data Supplementary Materials mmc2.


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nh?c one piece z

E-mail: chmhhv nus. Significant anion and solvent effects on the chemical shifts of the C2—H protons were found, which allows for a ranking of the anions in terms of their hydrogen-bond acceptor properties. Similar but less pronounced anion influences were detected for the 13 C C2 NMR resonances, while 1 J C2—H coupling constants appear to be anion and solvent independent. In particular, 1,3-diisopropylbenzimidazolium bromide A 14 is regularly used in our laboratories to prepare NHC complexes for donor strength determination using the HEP system. The preparation of salts B—G was most straightforward, and simply involved exposure of compound A to a slight excess of sodium or potassium salts containing the desired anion at ambient temperature AT.

This page has all of the required homework for the material covered in the first exam of the first semester of General Chemistry. Note: You are expected to go to the end of chapter problems in your textbook, find similar questions, and work out those problems as well.

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Microbial Cell Factories volume 18 , Article number: Cite this article. Metrics details. A Correction to this article was published on 02 January

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The transition elements or transition metals occupy the short columns in the center of the periodic table, between Group 2A and Group 3A. They are sometimes called the d -block elements , since in this region the d -orbitals are being filled in, and are also referred to as B-group elements since in most numbering systems of the columns on the periodic table the numerals of these groups are followed by the letter B. The period 7 transition metals are the naturally-occurring actinium Ac , and the artificially produced elements rutherfordium Rf , dubnium Db , seaborgium Sg , bohrium Bh , hassium Hs , meitnerium Mt , darmstadtium Ds , roentgenium Rg , and the as-yet unnamed ununbiium Uub. In the transition metals, the five d orbitals are being filled in, and the elements in general have electron configurations of n -1 d ns 2 , although there are some exceptions when electrons are shuffled around to produce half-filled or filled d subshells. All of the transition metals in their elemental forms are malleable and ductile except for mercury, which is a liquid at room temperature , and are good conductors of heat and electricity.

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