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1.
J Mass Spectrom ; 59(6): e5036, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38726689

ABSTRACT

Turmeric and ginger are extensively employed as functional ingredients due to their high content of curcuminoids and gingerols, considered the key bioactive compounds found in these roots. In this study, we present an innovative and fast method for the assay of curcuminoids and gingerols in different foods containing the two spices, with the aim of monitoring the quality of products from a nutraceutical perspective. The proposed approach is based on paper spray tandem mass spectrometry coupled with the use of a labeled internal standard, which has permitted to achieve the best results in terms of specificity and accuracy. All the calculated analytical parameters were satisfactory; accuracy values are around 100% for all spiked samples and the precision data result lower than 15%. The protocol was applied to several real samples, and to demonstrate its robustness and reliability, the results were compared to those arising from the common liquid chromatographic method.


Subject(s)
Curcuma , Fatty Alcohols , Tandem Mass Spectrometry , Zingiber officinale , Zingiber officinale/chemistry , Curcuma/chemistry , Tandem Mass Spectrometry/methods , Fatty Alcohols/analysis , Reproducibility of Results , Limit of Detection , Catechols/analysis , Food Analysis/methods , Curcumin/analysis , Curcumin/analogs & derivatives , Paper
2.
Ann Fam Med ; 22(3): 230-232, 2024.
Article in English | MEDLINE | ID: mdl-38806257

ABSTRACT

Reducing examination table paper (ETP) use may help curb carbon emissions from health care. Six participants applied Glo Germ (DMA International) to their hands before a common physical examination (abdominal, cardiorespiratory, hip and knee) both with and without ETP. After each exam, UV light was shined on the exam table and photographs were taken. The number of hand touches on ETP-covered areas and uncovered areas were tallied and compared using t tests. Despite covering more surface area, participants touched areas without ETP significantly more than ETP-covered areas (P <.05). Despite its continued use, patients do not have much hand contact with ETP during common clinical examinations.


Subject(s)
Family Practice , Paper , Physical Examination , Humans , Family Practice/methods , Physical Examination/methods , Female , Male , Adult , Ultraviolet Rays/adverse effects
3.
Compr Rev Food Sci Food Saf ; 23(3): e13373, 2024 May.
Article in English | MEDLINE | ID: mdl-38778547

ABSTRACT

The environmental challenges posed by plastic pollution have prompted the exploration of eco-friendly alternatives to disposable plastic packaging and utensils. Paper-based materials, derived from renewable resources such as wood pulp, non-wood pulp (bamboo pulp, straw pulp, reed pulp, etc.), and recycled paper fibers, are distinguished by their recyclability and biodegradability, making them promising substitutes in the field of plastic food packaging. Despite their merits, challenges like porosity, hydrophilicity, limited barrier properties, and a lack of functionality have restricted their packaging potential. To address these constraints, researchers have introduced antimicrobial agents, hydrophobic substances, and other functional components to improve both physical and functional properties. This enhancement has resulted in notable improvements in food preservation outcomes in real-world scenarios. This paper offers a comprehensive review of recent progress in hydrophobic antimicrobial paper-based materials. In addition to outlining the characteristics and functions of commonly used antimicrobial substances in food packaging, it consolidates the current research landscape and preparation techniques for hydrophobic paper. Furthermore, the paper explores the practical applications of hydrophobic antimicrobial paper-based materials in agricultural produce, meat, and seafood, as well as ready-to-eat food packaging. Finally, challenges in production, application, and recycling processes are outlined to ensure safety and efficacy, and prospects for the future development of antimicrobial hydrophobic paper-based materials are discussed. Overall, the emergence of hydrophobic antimicrobial paper-based materials stands out as a robust alternative to plastic food packaging, offering a compelling solution with superior food preservation capabilities. In the future, paper-based materials with antimicrobial and hydrophobic functionalities are expected to further enhance food safety as promising packaging materials.


Subject(s)
Anti-Infective Agents , Food Packaging , Hydrophobic and Hydrophilic Interactions , Paper , Food Packaging/methods , Anti-Infective Agents/chemistry , Anti-Infective Agents/pharmacology , Food Preservation/methods
4.
J Nanobiotechnology ; 22(1): 274, 2024 May 22.
Article in English | MEDLINE | ID: mdl-38773614

ABSTRACT

Small extracellular vesicle-derived microRNAs (sEV-miRNAs) have emerged as promising noninvasive biomarkers for early cancer diagnosis. Herein, we developed a molecular probe based on three-dimensional (3D) multiarmed DNA tetrahedral jumpers (mDNA-Js)-assisted DNAzyme activated by Na+, combined with a disposable paper-based electrode modified with a Zr-MOF-rGO-Au NP nanocomplex (ZrGA) to fabricate a novel biosensor for sEV-miRNAs Assay. Zr-MOF tightly wrapped by rGO was prepared via a one-step method, and it effectively aids electron transfer and maximizes the effective reaction area. In addition, the mechanically rigid, and nanoscale-addressable mDNA-Js assembled from the bottom up ensure the distance and orientation between fixed biological probes as well as avoid probe entanglement, considerably improving the efficiency of molecular hybridization. The fabricated bioplatform achieved the sensitive detection of sEV-miR-21 with a detection limit of 34.6 aM and a dynamic range from100 aM to 0.2 µM. In clinical blood sample tests, the proposed bioplatform showed results highly consistent with those of qRT-PCRs and the signal increased proportionally with the NSCLC staging. The proposed biosensor with a portable wireless USB-type analyzer is promising for the fast, easy, low-cost, and highly sensitive detection of various nucleic acids and their mutation derivatives, making it ideal for POC biosensing.


Subject(s)
Biosensing Techniques , Extracellular Vesicles , Limit of Detection , Metal-Organic Frameworks , MicroRNAs , Paper , Metal-Organic Frameworks/chemistry , Extracellular Vesicles/chemistry , Humans , Biosensing Techniques/methods , DNA, Catalytic/chemistry , Graphite/chemistry , Gold/chemistry , DNA/chemistry , Metal Nanoparticles/chemistry , Nucleic Acid Hybridization , Electrochemical Techniques/methods , Electrodes , Zirconium/chemistry
5.
Anal Chim Acta ; 1306: 342617, 2024 Jun 01.
Article in English | MEDLINE | ID: mdl-38692789

ABSTRACT

BACKGROUND: Alpha-fetoprotein (AFP) is a fetal protein that can indicate congenital anomalies such as Down syndrome and spinal canal blockage when detected at abnormal levels in pregnant women. Current AFP detection methods rely on invasive blood or serum samples, which require sophisticated equipment. From the many solutions proposed, colorimetric paper-based assays excel in point-of-care settings. The concept of paper-based ELISA (p-ELISA) enhances traditional methods, aligning with the ASSURED criteria for diagnostics in resource-limited regions. Despite success in microfluidic paper-based assay devices, laser printing remains underexplored for p-ELISA. Additionally, modifying the paper surface provides an additional layer of sensitivity enhancement. RESULTS: In this study, we developed a novel laser-printed paper-based ELISA (LP-pELISA) for rapid, sensitive, and noninvasive detection of AFP in saliva samples. The LP-pELISA platform was fabricated by printing hydrophobic barriers on filter paper using a laser printer, followed by depositing hydroxyapatite (HAp) as an immobilization material for the antibodies. The colorimetric detection was achieved using AuNPs functionalized with anti-AFP antibodies and silver nitrate enhancement. The LP-pELISA exhibited a linear response for AFP detection in both buffer and saliva samples over a range of 1.0-800 ng mL-1, with a limit of detection (LOD) reaching 1.0 ng mL-1. The assay also demonstrated good selectivity, repeatability, reproducibility, and stability. The LP-pELISA was further validated by testing spiked human saliva samples, showing its potential for point-of-care diagnosis of congenital disabilities. SIGNIFICANCE: The LP-pELISA is a noninvasive platform showcasing simplicity, cost-effectiveness, and user-friendliness, utilizing laser printing, hydroxyapatite modification, and saliva samples to efficiently detect AFP. Beyond its application for AFP, this method's versatility extends to other biomarkers, positioning it as a catalyst for the evolution of paper-based biosensors. The LP-pELISA holds promise as a transformative tool for point-of-care diagnostics, fostering advancements in healthcare with its innovative technology.


Subject(s)
Colorimetry , Durapatite , Enzyme-Linked Immunosorbent Assay , Lasers , Paper , Saliva , alpha-Fetoproteins , Humans , Saliva/chemistry , Durapatite/chemistry , alpha-Fetoproteins/analysis , Printing , Gold/chemistry , Limit of Detection , Antibodies, Immobilized/immunology , Antibodies, Immobilized/chemistry
6.
Anal Chim Acta ; 1308: 342575, 2024 Jun 15.
Article in English | MEDLINE | ID: mdl-38740448

ABSTRACT

BACKGROUND: Alzheimer's disease (AD) is a prevalent neurodegenerative disease with no effective treatment. Efficient and rapid detection plays a crucial role in mitigating and managing AD progression. Deep learning-assisted smartphone-based microfluidic paper analysis devices (µPADs) offer the advantages of low cost, good sensitivity, and rapid detection, providing a strategic pathway to address large-scale disease screening in resource-limited areas. However, existing smartphone-based detection platforms usually rely on large devices or cloud servers for data transfer and processing. Additionally, the implementation of automated colorimetric enzyme-linked immunoassay (c-ELISA) on µPADs can further facilitate the realization of smartphone µPADs platforms for efficient disease detection. RESULTS: This paper introduces a new deep learning-assisted offline smartphone platform for early AD screening, offering rapid disease detection in low-resource areas. The proposed platform features a simple mechanical rotating structure controlled by a smartphone, enabling fully automated c-ELISA on µPADs. Our platform successfully applied sandwich c-ELISA for detecting the ß-amyloid peptide 1-42 (Aß 1-42, a crucial AD biomarker) and demonstrated its efficacy in 38 artificial plasma samples (healthy: 19, unhealthy: 19, N = 6). Moreover, we employed the YOLOv5 deep learning model and achieved an impressive 97 % accuracy on a dataset of 1824 images, which is 10.16 % higher than the traditional method of curve-fitting results. The trained YOLOv5 model was seamlessly integrated into the smartphone using the NCNN (Tencent's Neural Network Inference Framework), enabling deep learning-assisted offline detection. A user-friendly smartphone application was developed to control the entire process, realizing a streamlined "samples in, answers out" approach. SIGNIFICANCE: This deep learning-assisted, low-cost, user-friendly, highly stable, and rapid-response automated offline smartphone-based detection platform represents a good advancement in point-of-care testing (POCT). Moreover, our platform provides a feasible approach for efficient AD detection by examining the level of Aß 1-42, particularly in areas with low resources and limited communication infrastructure.


Subject(s)
Alzheimer Disease , Amyloid beta-Peptides , Biomarkers , Enzyme-Linked Immunosorbent Assay , Paper , Smartphone , Alzheimer Disease/diagnosis , Alzheimer Disease/blood , Humans , Biomarkers/blood , Biomarkers/analysis , Amyloid beta-Peptides/analysis , Amyloid beta-Peptides/blood , Peptide Fragments/blood , Peptide Fragments/analysis , Lab-On-A-Chip Devices , Deep Learning , Automation , Microfluidic Analytical Techniques/instrumentation
7.
Anal Chim Acta ; 1308: 342639, 2024 Jun 15.
Article in English | MEDLINE | ID: mdl-38740452

ABSTRACT

BACKGROUND: Calcium and magnesium ions are highly abundant and important cations in human body. At the same time, both dyscalcemia and dysmagnesemia are frequently encountered in the clinical practice. As deficiency or excess of Ca(II) or Mg(II) can cause severe symptoms, determining these ions in serum is of great importance. Concentration of these ions in biological samples is typically assayed in clinical laboratories with the use of expensive and specialized equipment. Since those methods cannot be easily adapted for self-diagnosis purposes, there is a great need to develop a convenient tool for reliable determination of calcium and magnesium in serum at the point-of-care. RESULTS: The colorimetric methods employed for calcium and magnesium analysis were o-cresophtalein complexone assay and xylidyl blue assay, respectively. Analytical signal acquisition was accomplished using an ordinary flatbed scanner or smartphone and free software. For increased user-friendliness the device was optimized to perform simultaneous determination of calcium and magnesium ions in only 10 min. In the optimized conditions, the limit of detection for calcium ions was 0.09 mmol L-1, while for magnesium it was 0.04 mmol L-1. Determination of both ions requires only 4 µL of serum sample. The developed paper-based sensors were validated with control human serum samples and the obtained relative errors for majority of samples were below 20 %. SIGNIFICANCE: In this paper, a microfluidic paper-based analytical device for simultaneous determination of calcium and magnesium ions in human serum is reported for the first time. Additionally, this is also the first report on colorimetric determination in serum of any of these ions in paper-based format. Simultaneous detection of both ions allows for fast and user-friendly screening of disturbance in calcium and magnesium homeostasis.


Subject(s)
Calcium , Magnesium , Paper , Magnesium/blood , Humans , Calcium/blood , Colorimetry , Lab-On-A-Chip Devices , Microfluidic Analytical Techniques/instrumentation , Limit of Detection
8.
Anal Methods ; 16(19): 3007-3019, 2024 May 16.
Article in English | MEDLINE | ID: mdl-38695537

ABSTRACT

We present a colorimetric probe based on polyvinylpyrrolidone-capped gold nanoparticles (PVP-AuNPs) that is sensitive and selective for cysteine (Cys). A microfluidic paper-based analytical device (µ-PAD) with embedded dried PVP-AuNPs at the polyethersulfone (PES) paper surface is used for Cys detection. When thiol molecules attach to PVP-AuNPs in the presence of Cys, they clump together, and this causes the solution's color to shift from red to blue within 5 minutes. The device is capable of detecting Cys levels between 1.0 µM and 50.0 µM with a limit of detection (LOD) of 0.2 µM under optimized conditions. The stability of the µ-PAD was tested for 100 days, demonstrating re-dispersibility to detect Cys levels in blood. Dried PVP-AuNP-µPADs were integrated with blood plasma separation modules for point-of-care (POC) Cys detection. Consequently, the device shows potential as a self-sustaining, quantification platform with a recovery percentage ranging from 98.44 to 111.9 in clinical samples.


Subject(s)
Colorimetry , Cysteine , Gold , Limit of Detection , Metal Nanoparticles , Paper , Point-of-Care Systems , Gold/chemistry , Cysteine/blood , Cysteine/chemistry , Metal Nanoparticles/chemistry , Humans , Colorimetry/methods , Colorimetry/instrumentation , Povidone/chemistry , Microfluidic Analytical Techniques/instrumentation , Microfluidic Analytical Techniques/methods
9.
Anal Methods ; 16(19): 3131-3141, 2024 May 16.
Article in English | MEDLINE | ID: mdl-38712986

ABSTRACT

Plastics are ubiquitous in today's lifestyle, and their indiscriminate use has led to the accumulation of plastic waste in landfills and oceans. The waste accumulates and breaks into micro-particles that enter the food chain, causing severe threats to human health, wildlife, and the ecosystem. Environment-friendly and bio-based degradable materials offer a sustainable alternative to the vastly used synthetic materials. Here, a polylactic acid and carbon nanofiber-based membrane and a paper-based colorimetric sensor have been developed. The membrane had a surface area of 3.02 m2 g-1 and a pore size of 18.77 nm. The pores were evenly distributed with a pore volume of 0.0137 cm3 g-1. The membrane was evaluated in accordance with OECD guidelines and was found to be safe for tested aquatic and terrestrial models. The activated PLA-CNF membrane was further used as a bio-based electrode for the electrochemical detection of nitrates (NO3-) in water samples with a detection limit of 0.046 ppm and sensitivity of 1.69 × 10-4 A ppm-1 mm-2, whereas the developed paper-based colorimetric sensor had a detection limit of 156 ppm for NO3-. This study presents an environment-friendly, low-carbon footprint disposable material for sensing applications as a sustainable alternative to plastics.


Subject(s)
Carbon , Colorimetry , Nanofibers , Nitrates , Paper , Polyesters , Nanofibers/chemistry , Colorimetry/methods , Colorimetry/instrumentation , Nitrates/analysis , Nitrates/chemistry , Polyesters/chemistry , Carbon/chemistry , Electrochemical Techniques/methods , Electrochemical Techniques/instrumentation , Limit of Detection , Water Pollutants, Chemical/analysis , Electric Conductivity , Membranes, Artificial
10.
Molecules ; 29(10)2024 May 08.
Article in English | MEDLINE | ID: mdl-38792053

ABSTRACT

Sulfite, a widely used food additive, is subject to regulated labeling. The extraction of sulfite as the stable hydroxymethylsulfonate (HMS) form and its quantitative analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been recognized for their good sensitivity, selectivity, and versatility across various food materials. This study aimed to develop a cost-effective and simpler method for sulfite quantitation, while maintaining the superior sensitivity and selectivity of mass spectrometry (MS). To achieve this, we introduced paper spray ionization (PSI), an ambient desorption ionization technique that could achieve the direct measurement of analytes without employing separation. We also employed a novel internal standard (IS) structurally similar to the analyte, replacing the more expensive isotopically labeled IS. Although the PSI-MS/MS method developed in this study exhibited slightly lower analytical performance compared to the conventional LC-MS/MS, it remained effective for sulfite analysis in dried fruits.


Subject(s)
Fruit , Sulfites , Tandem Mass Spectrometry , Sulfites/analysis , Sulfites/chemistry , Tandem Mass Spectrometry/methods , Fruit/chemistry , Chromatography, Liquid/methods , Paper , Food Analysis/methods
11.
Sensors (Basel) ; 24(10)2024 May 12.
Article in English | MEDLINE | ID: mdl-38793927

ABSTRACT

Specialized cancer treatments have the potential to exploit glutamine dependence to increase patient survival rates. Glutamine diagnostics capable of tracking a patient's response to treatment would enable a personalized treatment dosage to optimize the tradeoff between treatment success and dangerous side effects. Current clinical glutamine testing requires sophisticated and expensive lab-based tests, which are not broadly available on a frequent, individualized basis. To address the need for a low-cost, portable glutamine diagnostic, this work engineers a cell-free glutamine biosensor to overcome assay background and signal-to-noise limitations evident in previously reported studies. The findings from this work culminate in the development of a shelf-stable, paper-based, colorimetric glutamine test with a high signal strength and a high signal-to-background ratio for dramatically improved signal resolution. While the engineered glutamine test is important progress towards improving the management of cancer and other health conditions, this work also expands the assay development field of the promising cell-free biosensing platform, which can facilitate the low-cost detection of a broad variety of target molecules with high clinical value.


Subject(s)
Biosensing Techniques , Glutamine , Metabolic Engineering , Biosensing Techniques/methods , Glutamine/metabolism , Metabolic Engineering/methods , Humans , Genetic Engineering/methods , Paper , Colorimetry/methods , Cell-Free System
12.
J Hazard Mater ; 472: 134419, 2024 Jul 05.
Article in English | MEDLINE | ID: mdl-38691993

ABSTRACT

The contamination of paper products by various chemicals has been reported on a global level, but to date, no published research has investigated pharmaceutical contamination of paper-based products. In this study, pharmaceutical analysis was conducted on 42 samples collected from various points of the recycled paper value chain in Cape Town, South Africa, which included the various grades that may be included in the manufacturing of recycled paperboard. The analysis was achieved by ultrasonic-assisted extraction of paper samples before detection by UHPLC-Q Orbitrap. Quantification limits ranged from 1.15 pg/g for ketoprofen to 46.07 pg/g for methocarbamol. Pharmaceuticals identified in newspaper samples were dexamethasone, ketoprofen, and 17ß-estradiol. The latter was also detected in paper shopping bags (up to 697.49 ng/g), infant bathtub packaging (280.62 ng/g), battery packaging (137.43 ng/g), and an egg carton (170.47 ng/g). Carbamazepine was also prominent with its concentration reaching 13.02 ng/g in a vegetable box. Suspect screening tentatively identified 14 additional pharmaceuticals in paper samples, with minocycline, prazepam, and anabolic steroids appearing more prominently. This pioneering study indicated that unintentional pharmaceutical exposure had expanded beyond environmental media to consumer products.


Subject(s)
Paper , Recycling , Pharmaceutical Preparations/analysis , South Africa , Chromatography, High Pressure Liquid
13.
Protein Expr Purif ; 220: 106490, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38697589

ABSTRACT

The production of fermentable sugars from lignocellulosic biomass is achieved by the synergistic action of a group of enzymes called cellulases. Cellulose is a long chain of chemically linked glucoses by ß-1,4 bonds. The enzyme ß-1,4-endoglucanase is the first cellulase involved in the degradation, breaking the bond of the amorphous regions. A ß-1,4-endoglucanase enzyme with high activity was obtained from a Bacillus subtilis strain isolated from wastewater of a pulp and paper mill. Sequencing and bioinformatic analysis showed that the gene amplified by PCR consisting of 1407 nucleotides and coding for a ß-1,4-endoglucanase enzyme of approximately 55 kDa. The open reading frame (ORF) encoding the mature endoglucanase (eglS) was successfully inserted in a modified cloning plasmid (pITD03) and into the pYD1 plasmid used for its expression in yeast. Carboxymethylcellulose (CMC) plate assay, SDS-PAGE, and zymogram confirmed the production and secretion by the transformed E. coli BL21-SI strain of a 39 kDa ß-1,4-endoglucanase consistent with the catalytic domain without the cellulose-binding module (CBM). The results showed that the truncated ß-1,4-endoglucanase had higher activity and stability.


Subject(s)
Bacillus subtilis , Cellulase , Paper , Recombinant Proteins , Wastewater , Bacillus subtilis/genetics , Bacillus subtilis/enzymology , Bacillus subtilis/isolation & purification , Wastewater/microbiology , Wastewater/chemistry , Cellulase/genetics , Cellulase/chemistry , Cellulase/biosynthesis , Cellulase/isolation & purification , Cellulase/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/chemistry , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , Recombinant Proteins/biosynthesis , Bacterial Proteins/genetics , Bacterial Proteins/chemistry , Bacterial Proteins/isolation & purification , Bacterial Proteins/biosynthesis , Bacterial Proteins/metabolism , Escherichia coli/genetics , Escherichia coli/metabolism , Cloning, Molecular , Gene Expression
14.
Cancer Med ; 13(9): e7189, 2024 May.
Article in English | MEDLINE | ID: mdl-38706442

ABSTRACT

OBJECTIVES: Endoscopic ultrasound-guided tissue acquisition (EUS-TA) is used for pathological diagnosis and obtaining samples for molecular testing, facilitating the initiation of targeted therapies in patients with pancreatic cancer. However, samples obtained via EUS-TA are often insufficient, requiring more efforts to improve sampling adequacy for molecular testing. Therefore, this study investigated the use of oil blotting paper for formalin fixation of samples obtained via EUS-TA. METHODS: This prospective study enrolled 42 patients who underwent EUS-TA for pancreatic cancer between September 2020 and February 2022 at the Osaka International Cancer Institute. After a portion of each sample obtained via EUS-TA was separated for routine histological evaluation, the residual samples were divided into filter paper and oil blotting paper groups for analysis. Accordingly, filter paper and oil blotting paper were used for the formalin fixation process. The total tissue, nuclear, and cytoplasm areas of each sample were quantitatively evaluated using virtual slides, and the specimen volume and histological diagnosis of each sample were evaluated by an expert pathologist. RESULTS: All cases were cytologically diagnosed as adenocarcinoma. The area ratios of the total tissue, nuclear, and cytoplasmic portions were significantly larger in the oil blotting paper group than in the filter paper group. The frequency of cases with large amount of tumor cells was significantly higher in the oil blotting paper group (33.3%) than in the filter paper group (11.9%) (p = 0.035). CONCLUSIONS: Oil blotting paper can increase the sample volume obtained via EUS-TA on glass slides and improve sampling adequacy for molecular testing.


Subject(s)
Formaldehyde , Pancreatic Neoplasms , Tissue Fixation , Humans , Pancreatic Neoplasms/pathology , Pancreatic Neoplasms/diagnostic imaging , Prospective Studies , Male , Female , Tissue Fixation/methods , Aged , Middle Aged , Endosonography/methods , Specimen Handling/methods , Adenocarcinoma/pathology , Adenocarcinoma/diagnostic imaging , Aged, 80 and over , Paper , Endoscopic Ultrasound-Guided Fine Needle Aspiration/methods
15.
Biosens Bioelectron ; 257: 116292, 2024 Aug 01.
Article in English | MEDLINE | ID: mdl-38653014

ABSTRACT

We report the development and initial validation of a paper-based nucleic acid testing platform that integrates Loop-mediated isothermal amplification (LAMP) with clustered regularly interspaced short palindromic repeats (CRISPR) technology, referred to as PLACID (Paper-based LAMP-CRISPR Integrated Diagnostics). LAMP eliminates the need for thermal cycling, resulting in simplified instrumentation, and the CRISPR-associated protein (Cas 12a) system eliminates false positive signals from LAMP products, resulting in highly selective and sensitive assays. We optimized the assay to perform both amplification and detection entirely on paper, eliminating the need for complex fluid handling steps and lateral flow assay transfers. Additionally, we engineered a smartphone-operated system that includes a low-powered, non-contact IR heating chamber to actuate paper-based LAMP and CRISPR reactions and enable the detection of fluorescent signals from the paper. The platform demonstrates high specificity and sensitivity in detecting nucleic acid targets with a limit of detection of 50 copies/µL. We integrate an equipment-free sample preparation separation technology designed to streamline the preparation of crude samples prior to nucleic acid testing. The practical utility of our platform is demonstrated by the successful detection of spiked SARS-CoV-2 RNA fragments in saliva, E. Coli in soil, and pathogenic E. Coli in clinically fecal samples of infected patients. Furthermore, we demonstrate that the paper-based LAMP CRISPR chips employed in our assays possess a shelf life of several weeks, establishing them as viable candidates for on-site diagnostics.


Subject(s)
Biosensing Techniques , COVID-19 , CRISPR-Cas Systems , Molecular Diagnostic Techniques , Nucleic Acid Amplification Techniques , Paper , SARS-CoV-2 , Nucleic Acid Amplification Techniques/methods , Nucleic Acid Amplification Techniques/instrumentation , Humans , Biosensing Techniques/methods , SARS-CoV-2/genetics , SARS-CoV-2/isolation & purification , COVID-19/diagnosis , COVID-19/virology , Molecular Diagnostic Techniques/methods , Molecular Diagnostic Techniques/instrumentation , CRISPR-Cas Systems/genetics , Limit of Detection , Clustered Regularly Interspaced Short Palindromic Repeats/genetics , Equipment Design , COVID-19 Nucleic Acid Testing/methods , COVID-19 Nucleic Acid Testing/instrumentation , Escherichia coli/genetics , Escherichia coli/isolation & purification , CRISPR-Associated Proteins/genetics , Smartphone
16.
Biosens Bioelectron ; 257: 116300, 2024 Aug 01.
Article in English | MEDLINE | ID: mdl-38657378

ABSTRACT

Developing simple, inexpensive, fast, sensitive, and specific probes for antibiotic-resistant bacteria is crucial for the management of urinary tract infections (UTIs). We here propose a paper-based sensor for the rapid detection of ß-lactamase-producing bacteria in the urine samples of UTI patients. By conjugating a strongly electronegative group -N+(CH3)3 with the core structures of cephalosporin and carbapenem antibiotics, two visual probes were achieved to respectively target the extended-spectrum/AmpC ß-lactamases (ESBL/AmpC) and carbapenemase, the two most prevalent factors causing antibiotic resistance. By integrating these probes into a portable paper sensor, we confirmed 10 and 8 cases out of 30 clinical urine samples as ESBL/AmpC- and carbapenemase-positive, respectively, demonstrating 100% clinical sensitivity and specificity. This paper sensor can be easily conducted on-site, without resorting to bacterial culture, providing a solution to the challenge of rapid detection of ß-lactamase-producing bacteria, particularly in resource-limited settings.


Subject(s)
Biosensing Techniques , Paper , Urinary Tract Infections , beta-Lactamases , beta-Lactamases/metabolism , beta-Lactamases/chemistry , Humans , Urinary Tract Infections/microbiology , Urinary Tract Infections/diagnosis , Biosensing Techniques/methods , Anti-Bacterial Agents/therapeutic use , Anti-Bacterial Agents/pharmacology , Bacterial Proteins , Bacteria/isolation & purification , Bacteria/enzymology , Cephalosporins/chemistry , Carbapenems/pharmacology
17.
Int J Biol Macromol ; 268(Pt 2): 131790, 2024 May.
Article in English | MEDLINE | ID: mdl-38677693

ABSTRACT

The demand for paper-based packaging materials as an alternative to incumbent disposable petroleum-derived polymers for food packaging applications is ever-growing. However, typical paper-based formats are not suitable for use in unconventional applications due to inherent limitations (e.g., excessive hydrophilicity, lack antimicrobial ability), and accordingly, enabling new capabilities is necessity. Herein, a simple and environmentally friendly strategy was proposed to introduce antimicrobial and hydrophobic functions to cellulose paper through successive chemical grafting of 3-aminopropyltriethoxysilane (APS) and cinnamaldehyde (CA). The results revealed that cellulose paper not only showed long-term antibacterial effect on different bacteria, but also inhibited a wide range of fungi. Encouragingly, the modified paper, which is fluorine-free, displays a high contact angle of 119.7°. Thus, even in the wet state, the modified paper can still maintain good mechanical strength. Meanwhile, the multifunctional composite papers have excellent biocompatibility and biodegradability. Compared with ordinary cellulose paper, multifunctional composite paper can effectively prolong the shelf life of strawberries. Therefore, the multifunctional composite paper represents good application potential as a fruit packaging material.


Subject(s)
Acrolein , Cellulose , Food Packaging , Fragaria , Hydrophobic and Hydrophilic Interactions , Paper , Cellulose/chemistry , Cellulose/analogs & derivatives , Acrolein/analogs & derivatives , Acrolein/chemistry , Acrolein/pharmacology , Fragaria/microbiology , Food Packaging/methods , Anti-Infective Agents/pharmacology , Anti-Infective Agents/chemistry , Silanes/chemistry , Food Preservation/methods , Propylamines/chemistry , Microbial Sensitivity Tests
18.
Talanta ; 274: 126053, 2024 Jul 01.
Article in English | MEDLINE | ID: mdl-38599121

ABSTRACT

Borax is strictly regulated in the food processing and pharmaceutical industry due to its physiological toxicity, and the development of a direct analytical method is essential for effectively monitoring the borax abuse. In this work, the fluorescence properties of flavonoids, including flavones, isoflavones and flavonols, were systematically investigated from aqueous to borax solutions, and it was found that the weak intrinsic fluorescence of flavonols could be pervasively sensitized by borax. A natural flavonol, morin, was subsequently chosen as a representative probe to develop a turn-on fluorescence sensing method for borax analysis, which achieved a linear response spanning four orders of magnitude with a detection limit of 1.07 µM (0.22 µg mL-1 in terms of Na2B4O7 content). Furthermore, a smartphone-assisted paper-based test device was designed and constructed by 3D printing technology. Using morin-impregnated test strips as the carrier, the borax could be visually detected by the RGB signals of the captured images, with a detection limit of 0.13 mM (27.05 µg mL-1 for Na2B4O7). Combining ion exchange treatment for food samples and sodium periodate oxidation for drug samples, the developed methods were successfully applied for the direct analysis of borax in various products with the recoveries of 86.9-106.3% for traditional fluorescence analysis and 82.7-108.8% for smartphone-assisted fluorescence sensing. The fluorescence property of the morin-borax system was studied using time-dependent density functional theory, and the sensing mechanism was discussed in conjunction with experimental research.


Subject(s)
Flavones , Flavonoids , Flavonols , Paper , Smartphone , Spectrometry, Fluorescence , Flavonols/analysis , Spectrometry, Fluorescence/methods , Flavonoids/analysis , Borates/chemistry , Limit of Detection , Fluorescent Dyes/chemistry , Fluorescence
19.
Lab Chip ; 24(9): 2454-2467, 2024 Apr 30.
Article in English | MEDLINE | ID: mdl-38644805

ABSTRACT

Safe, accurate, and reliable analysis of urinary biomarkers is clinically important for early detection and monitoring of the progression of chronic kidney disease (CKD), as it has become one of the world's most prevalent non-communicable diseases. However, current technologies for measuring urinary biomarkers are either time-consuming and limited to well-equipped hospitals or lack the necessary sensitivity for quantitative analysis and post a health risk to frontline practitioners. Here we report a robust paper-based dual functional biosensor, which is integrated with the clinical urine sampling vial, for the simultaneous and quantitative analysis of pH and glucose in urine. The pH sensor was fabricated by electrochemically depositing IrOx onto a paper substrate using optimised parameters, which enabled an ultrahigh sensitivity of 71.58 mV pH-1. Glucose oxidase (GOx) was used in combination with an electrochemically deposited Prussian blue layer for the detection of glucose, and its performance was enhanced by gold nanoparticles (AuNPs), chitosan, and graphite composites, achieving a sensitivity of 1.5 µA mM-1. This dual function biosensor was validated using clinical urine samples, where a correlation coefficient of 0.96 for pH and 0.98 for glucose detection was achieved with commercial methods as references. More importantly, the urine sampling vial was kept sealed throughout the sample-to-result process, which minimised the health risk to frontline practitioners and simplified the diagnostic procedures. This diagnostic platform, therefore, holds high promise as a rapid, accurate, safe, and user-friendly point-of-care (POC) technology for the analysis of urinary biomarkers in frontline clinical settings.


Subject(s)
Biosensing Techniques , Paper , Point-of-Care Systems , Humans , Hydrogen-Ion Concentration , Gold/chemistry , Glucose/analysis , Urinalysis/instrumentation , Glucose Oxidase/chemistry , Glucose Oxidase/metabolism , Electrochemical Techniques , Metal Nanoparticles/chemistry , Graphite/chemistry , Biomarkers/urine
20.
Biosensors (Basel) ; 14(4)2024 Apr 03.
Article in English | MEDLINE | ID: mdl-38667165

ABSTRACT

The exploration into nanomaterial-based nonenzymatic biosensors with superb performance in terms of good sensitivity and anti-interference ability in disease marker monitoring has always attained undoubted priority in sensing systems. In this work, we report the design and synthesis of a highly active nanocatalyst, i.e., palladium and platinum nanoparticles (Pt&Pd-NPs) decorated ultrathin nanoporous gold (NPG) film, which is modified on a homemade graphene paper (GP) to develop a high-performance freestanding and flexible nanohybrid electrode. Owing to the structural characteristics the robust GP electrode substrate, and high electrochemically catalytic activities and durability of the permeable NPG support and ultrafine and high-density Pt&Pd-NPs on it, the resultant Pt&Pd-NPs-NPG/GP electrode exhibits excellent sensing performance of low detection limitation, high sensitivity and anti-interference capability, good reproducibility and long-term stability for the detection of small molecular biomarkers hydrogen peroxide (H2O2) and glucose (Glu), and has been applied to the monitoring of H2O2 in different types of live cells and Glu in body fluids such as urine and fingertip blood, which is of great significance for the clinical diagnosis and prognosis in point-of-care testing.


Subject(s)
Biomarkers , Biosensing Techniques , Electrochemical Techniques , Gold , Graphite , Metal Nanoparticles , Palladium , Platinum , Graphite/chemistry , Gold/chemistry , Platinum/chemistry , Palladium/chemistry , Metal Nanoparticles/chemistry , Biomarkers/urine , Humans , Hydrogen Peroxide , Alloys/chemistry , Glucose/analysis , Electrodes , Paper
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