Paper-Based Test for Rapid On-Site Screening of SARS-CoV-2 in Clinical Samples

Detection methods for monitoring infectious pathogens has never been more important given the need to contain the spread of the COVID-19 pandemic. Herein we propose a highly sensitive magnetic-focus-enhanced lateral flow assay (mLFA) for the detection of SARS-CoV-2. The proposed mLFA is simple and requires only lateral flow strips and a reusable magnet to detect very low concentrations of the virus particles. The magnetic focus enhancement is achieved by focusing the SARS-CoV-2 conjugated magnetic probes in the sample placed in the lateral flow (LF) strips for improved capture efficiency, while horseradish peroxidase (HRP) was used to catalyze the colorimetric reaction for the amplification of the colorimetric signal. With the magnetic focus enhancement and HRP-based amplification, the mLFA could yield a highly sensitive technology for the recognition of SARS-CoV-2. The developed methods could detect as low as 400 PFU/mL of SARS-CoV-2 in PBS buffer based on the visible blue dots on the LF strips. The mLFA could recognize 1200 PFU/mL of SARS-CoV-2 in saliva samples. With clinical nasal swab samples, the proposed mLFA could achieve 66.7% sensitivity and 100% specificity.

A Convenient and Label-Free Colorimetric Detection for L-Histidine Based on Inhibition of Oxidation of 3,3′,5,5′-Tetramethylbenzidine-H2O2 System Triggered by Copper Ions

L-Histidine (L-His) is an essential amino acid, which is used to synthesize proteins and enzymes. The concentration of L-His in the body is controlled to regulate tissue growth and repair of tissues. In this study, a rapid and sensitive method was developed for colorimetric L-his detection using Cu2+ ions to inhibit the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB)–H2O2 system. H2O2 can oxidize TMB to oxTMB in the presence of copper, and the change in color from colorless (TMB) to blue (oxTMB) is similar to that observed in the presence of peroxidase. However, because the imidazole ring and carboxyl group of L-His can coordinate with Cu2+ ions to form stable L-His–Cu2+ complexes, the color of the TMB–H2O2 solution remains unchanged after the addition of L-His. Therefore, because L-His effectively hinders the colorimetric reaction of TMB with H2O2, this assay can be used to quantitatively determine the concentration of L-His in samples. Under optimized conditions, our colorimetric sensor exhibited two linear ranges of 60 nM to 1 μM and 1 μM to 1 mM for L-His detection and a detection limit of 50 nM (S/N = 3); furthermore, the assay can be performed within 20 min. Moreover, the proposed assay was used to determine the concentration of L-His in urine samples, suggesting that this convenient and label-free colorimetric method presents promising applications in bioanalytical chemistry and clinical diagnosis.

Colorimetric Immunological Paper-based Assay for Exosome Detection

This paper reports the development of colorimetric immunological paper-based assay for exosome detection. The paper-based device was fabricated with lamination technique for easy handling and create hydrophilic/hydrophobic region for analytical paper-based devices. Exosome-specific antibody was coated onto the paper-based devices as a biosensing platform to detect exosome sample from the cell culture media. This assay employed a colorimetric reaction which is followed by reaction between horseradish peroxidase (HRP) and 3,3’,5,5’-tetramethylbenzidine substrate (TMB). The colorimetric readout was qualitatively evaluated by naked eyes and was quantitatively assessed by image processing software. The result indicated that this assay faces many challenges. First, the exosome concentration may be inadequate to reach detectable range. Second, high background signal due to non-specific binding on the platform results in lack of sensitivity for exosome detection. Therefore, modification on the paper should promote protein binding for specific target and prevent non-specific binding to reduce the high background signal.

Automated ELISA on-chip for the detection of an-ti-SARS-CoV-2 antibodies

The COVID-19 pandemic has been the most critical public health issue in modern history due to its highly infectious and deathly potential; and the limited access to massive, low-cost, and relia-ble testing has significantly worsened the crisis. The recovery and the vaccination of millions of people against COVID-19, have made serological tests highly relevant to identify the presence and levels of SARS-CoV-2 antibodies. Due to its advantages, microfluidic-based technologies repre-sent an attractive alternative to the conventional testing methodologies used for these purposes. In this work, we describe the development of an automated ELISA on-chip capable of detecting an-ti-SARS-CoV-2 antibodies in serum samples from COVID-19 patients and vaccinated individu-als. The colorimetric reactions were analyzed with a microplate reader. No statistically signifi-cant differences were observed when comparing the results of our automated ELISA on-chip against the ones obtained from a traditional ELISA on a microplate. Moreover, we demonstrated that it is possible to carry out the analysis of the colorimetric reaction by performing basic image analysis of photos taken with a smartphone, which constitutes a useful alternative when lacking specialized equipment or a laboratory setting. Our automated ELISA on-chip has the potential to be used in a clinical setting and mitigate some of the burden caused by testing deficiencies

NQS-Doped PDMS Solid Sensor: From Water Matrix to Urine Enzymatic Application

The development of in situ analytical devices has gained outstanding scientific interest. A solid sensing membrane composed of 1,2-naphthoquinone-4-sulfonate (NQS) derivatizing reagent embedded into a polymeric polydimethylsiloxane (PDMS) composite was proposed for in situ ammonium (NH4+) and urea (NH2CONH2) analysis in water and urine samples, respectively. Satisfactory strategies were also applied for urease-catalyzed hydrolysis of urea, either in solution or glass-supported urease immobilization. Using diffuse reflectance measurements combined with digital image processing of color intensity (RGB coordinates), qualitative and quantitative analyte detection was assessed after the colorimetric reaction took place inside the sensing membrane. A suitable linear relationship was found between the sensor response and analyte concentration, and the results were validated by a thymol-PDMS-based sensor based on the Berthelot reaction. The suggested sensing device offers advantages such as rapidity, versatility, portability, and employment of non-toxic reagents that facilitate in situ analysis in an energy-efficient manner.

Multiple SERS Detection of Phenol Derivatives in Tap Water

Phenol and some of its derivatives are products of the petrochemical industry. These compounds are characterized by their exceptional ability to persist in media and reach both food and water used by human beings. The consumption of these compounds has harmful effects on health, producing both acute and chronic effects—among the most prominent damages are teratogenicity, mutagenicity and carcinogenicity. Compounds such as phenol, ortho-cresol, and 1-napthol are listed by agencies such as the EPA. The aim of this work was the development of a rapid method for the simultaneous detection of these compounds in water samples. The method was based on the colorimetric reaction between phenol derivatives and Gibbs reagent, which forms indophenolates; using increased surface Raman spectroscopy together with statistical methods, SERS spectra were acquired, which were then analyzed. The developed method allows one to at least equalize the detection limits of the colorimetric method through UV–VIS spectroscopy and to discriminate among the three mixed phenols in at least binary aqueous samples. The major advantage of the method is the possibility of discriminating between phenol spectra quickly and easily.

Colorimetric Detection of Hg2+ Based on the Promotion of Oxidase-Like Catalytic Activity of Ag Nanowires

A portable Hg[Formula: see text]nanosensor was developed based on the colorimetric reaction by using the unmodified Ag nanowires (Ag NWs). Ag NWs were synthesized by a solvothermal method, with the length longer than 20[Formula: see text][Formula: see text]m and the diameter of [Formula: see text][Formula: see text]nm. The colorimetric assay can be affected by pH, temperature and the amount of Ag NWs, with the optimum parameters being 5, [Formula: see text]C and 100[Formula: see text][Formula: see text]L, respectively. The developed nanosensor presents excellent selectivity for Hg[Formula: see text]. The dynamic detection range is 25[Formula: see text]5000 ppb, and the limit of detection (LOD) for Hg[Formula: see text] is 19.9[Formula: see text]ppb. The developed Hg[Formula: see text] sensor shows great potentials in environmental monitoring and onsite analysis of Hg[Formula: see text].

Development of test placers for blister colorimetric analysis of phosphate ions

Variants of the compositions of a series of placers for visually blister colorimetric determination of phosphate ion concentrations in a blister cell (tablet for tablets) containing a mixture of reagents are proposed. The placers allow quantitative determination of the phosphate concentration in various objects without preliminary preparation of reagent solutions. The application of reagents to the surface of silica gel makes it possible to isolate all reagents until a chemical reaction occurs. The composition and conditions of deposition of crystalline substances – reagents capable of providing a sufficiently high acidity with the addition of a few drops of a solution and an optimal reducing agent for blister conditions were optimized. The visual colorimetric reaction is carried out under conditions when the volume of the added solution is 0.1–0.2 ml.

Colorimetric detection of glyphosate: towards a handmade and portable analyzer

Glyphosate (GFT) is a widely used herbicide, considered toxic and a probable carcinogen. The main challenge is its detection, usually requiring expensive and laborious methodologies. Herein, we report a colorimetric detection of GFT, using a derivatization reaction with 2,4-dinitrofluorobenzene (DNFB) that leads to a yellow-colored product. This is undertaken under mild conditions (weakly basic aqueous medium and ambient conditions). A thorough kinetic study was carried out, showing that the derivatization reaction with GFT predominates over the hydrolysis of DNFB. Hence, the colorimetric product is the major product formed, which was fully characterized by nuclear magnetic resonance. Finally, a portable, handmade and cheap colorimeter was used to detect and quantify GFT, relying on the colorimetric reaction proposed. Simulating real contaminated samples, it was possible to analyze in just 10 min, with less than 7 % of error of the nominal concentration. Overall, a highly sustainable approach is shown for an herbicide monitoring, with a simple and mild derivatization reaction that does not require purification and leads to a colorimetric product. Moreover, a simple apparatus with low time analysis is proposed that uses a problematic electronic trash: cellphone chargers. This cheapens the process and allows field analysis that can be extended to other agrochemicals.