Search Results (20)

Point-of-care (POC) antigen detection plays a crucial role in curbing the spread of viruses. Paper-based fluorescence aptasensors are expected to offer a low-cost tool to meet the needs of decentralized POC diagnosis. Herein, we report on a fluorescent paper-based sensing system for detecting the SARS-CoV-2 spike protein. The sensing system was constructed by loading multi-layer Nb₂C MXene nano-quenchers and carbon-dot-labeled aptamer (G-CDs@Apt) probes onto a mixed cellulose ester (MCE) paper substrate. On the Nb₂C MXene/G-CDs@Apt sensing paper, abundant G-CDs@Apt probes were attached to the multilayer MXene nano-quenchers and kept in a fluorescence-off state, while recognition of the target detached the G-CDs@Apt probes formed the nano--quenchers, resulting in fluorescence recovery of the sensing paper. The developed paper-based sensor performed well in the one-step detection of the SARS-CoV-2 S1 protein with a detection limit of 0.067 ng/mL (0.335 pg/test). The assay exhibited good selectivity and anti-interference in the detection of the SARS-CoV-2 S1 protein in artificial saliva. Moreover, the paper-based aptasensor was successfully used to detect the SARS-CoV-2 S1 protein in actual environmental samples with recoveries of 90.87–100.55% and relative standard deviations of 1.52–3.41%. The proposed technology provides a cost-effective alternative to traditional antibody test strips for a wide range of POC diagnostic applications. Full article

Lateral flow immunoassay (LFIA) technology serves a significant role as a simple and rapid biosensor in the detection of influenza viruses. The focus of this study is the development of a rapid and convenient screening method for influenza B virus (IBV) proteins using a fluorescence lateral flow biosensor based on Ag-doped ZnIn₂S₄ quantum dots (Ag: ZIS QDs) as signal reporters. These Ag: ZIS QDs-emitting orange fluorescence are loaded onto dendritic mesoporous silica nanoparticles (DMSNs) and are further coated with a layer of silica shell to form a core–shell structured composite nanomaterial (SiO₂ @ Ag: ZIS QDs @ DMSNs). The orange fluorescence effectively eliminates the interference of blue background fluorescence, significantly enhancing the detection sensitivity. This technology demonstrates outstanding performance in the immediate detection of IBV, with a minimum detection limit of 1 ng/mL, compared to the traditional colloidal gold strip with a detection limit of 6 ng/mL. Furthermore, both intra-assay and inter-assay coefficients of variation (CV) are less than 9%. This method holds promise for wide application in early diagnosis, epidemiological investigation, and epidemic surveillance of IBV. Full article

The sensitivity of fluorescent lateral flow immunoassay (LFIA) test strips is compromised by the low fluorescence intensity of the signaling molecules. In this study, we synthesized novel phosphorus-doped carbon-dot-based dendritic mesoporous silica nanoparticles (DMSNs-BCDs) with a quantum yield as high as 93.7% to break this bottleneck. Meanwhile, the in situ growth method increased the loading capacity of carbon dots on dendritic mesoporous silica, effectively enhancing the fluorescence intensity of the composite nanospheres. Applied DMSNs-BCDs in LFIA can not only semi-quantitatively detect a single component in a short time frame (procalcitonin (PCT), within 15 min) but also detect the dual components with a low limit of detection (LOD) (carbohydrate antigen 199 (CA199) LOD: 1 U/mL; alpha-fetoprotein (AFP) LOD: 0.01 ng/mL). And the LOD of PCT detection (0.01 ng/mL) is lower by 1.7 orders of magnitude compared to conventional colloidal gold strips. For CA199, the LOD is reduced by a factor of four compared to LFIA using gold nanoparticles as substrates, and for AFP, the LOD is lowered by two orders of magnitude compared to colloidal gold LFIA. Furthermore, the coefficients of variation (CV) for intra-assay and inter-assay measurements are both less than 11%. Full article

Acinetobacter baumannii (A. baumannii) is among the main pathogens that cause nosocomial infections. The ability to rapidly and accurately detect A. baumannii and its drug resistance is essential for blocking secondary infections and guiding treatments. In this study, we reported a nucleic acid fluorescent lateral flow assay (NFLFA) to identify A. baumannii and carbapenem-resistant A. baumannii (CRAB) in a rapid and quantitative manner by integrating loop-mediated isothermal amplification (LAMP) and silica–based multilayered quantum dot nanobead tag (Si@MQB). First, a rapid LAMP system was established and optimised to support the effective amplification of two bacterial genes in 35 min. Then, the antibody-modified Si@MQB was introduced to capture the two kinds of amplified DNA sequences and simultaneously detect them on two test lines of a LFA strip, which greatly improved the detection sensitivity and stability of the commonly used AuNP-based nucleic acid LFA. With these strategies, the established LAMP-NFLFA achieved detection limits of 199 CFU/mL and 287 CFU/mL for the RecA (house-keeping gene) and bla_OXA-23 (drug resistance gene) genes, respectively, within 43 min. Furthermore, the assay exhibited good repeatability and specificity for detecting target pathogens in real complex specimens and environments; thus, the proposed assay undoubtedly provides a promising and low-cost tool for the on-site monitoring of nosocomial infections. Full article

Salmonella is one of the leading causes of foodborne illnesses worldwide, with poultry products being a major source of contamination. Thus, the detection of salmonella in commercial poultry products is crucial to minimize the effects on public health. Electrochemical sensors are promising tools for bacteria detection due to their sensitivity, simplicity, and potential for on-site analysis. In this work, a three-dimensional (3D) printed electrochemical immunosensor for the determination of Salmonella typhimurium in fresh chicken through a sandwich immunoassay employing biotinylated anti-S. typhimurium antibody followed by streptavidin labeled with Cd/Se ZnS quantum dots (QDs) is presented. The device features three carbon-black polylactic acid electrodes and a holder, and the quantification of S. typhimurium is performed by anodic stripping voltametric (ASV) determination of the Cd(II) released after acidic dissolution of the QDs. To enhance sensitivity, an electroplated bismuth film was deposited on the working electrode, achieving a detection limit of 5 cfu/mL in a total assay time of 25 min, whereas 5 h of sample pre-enrichment was required for the detection of 1 cfu/25 mL of chicken rinse and chicken broth. The method is accurate, with %recovery values ranging from 93.3 to 113% in fresh chicken samples, and repeatable with intra- and inter- assay coefficient of variations <2 and 5%, respectively, indicating the suitability of the proposed immunosensor for the detection of S. typhimurium at the point-of-need. Full article

Maize chlorotic mottle virus (MCMV) is the only species in the Mahromovirus genus and is often co-infected with one or several viruses of the Potyvirus genus, posing a great threat to the global maize industry. Effective viral integrated management measures are dependent on the timely and proper detection of the causal agent of the disease. In this work, six super-sensitive and specific monoclonal antibodies (mAbs) against MCMV were first prepared using purified MCMV virions as the immunogen. Then, the Dot enzyme-linked immunosorbent assay (Dot-ELISA) was established based on the obtained mAbs, and it can detect MCMV in infected maize leaf crude extracts diluted up to 1:10,240-fold (w/v, g/mL). Furthermore, a rapid and user-friendly Au nanoparticle-based immunochromatographic test strip (AuNP-ICTS) based on paired mAbs 7B12 and 17C4 was created for monitoring MCMV in point-of-care tests, and it can detect the virus in a 25,600-fold dilution (w/v, g/mL) of MCMV-infected maize leaf crude extracts. The whole test process for ICTS was completed in 10 min. Compared with conventional reverse transcription-polymerase chain reaction (RT-PCR), the detection endpoint of both serological methods is higher than that of RT-PCR, especially the Dot-ELISA, which is 12.1 times more sensitive than that of RT-PCR. In addition, the detection results of 20 blinded maize samples by the two serological assays were consistent with those of RT-PCR. Therefore, the newly created Dot-ELISA and AuNP-ICTS exhibit favorable application potential for the detection of MCMV in plant samples. Full article

Plum pox virus (PPV) is a causal agent of the stone fruit tree sharka disease that often causes enormous economic losses. Due to its worldwide distribution and economic importance, rapid and reliable diagnostic technologies are becoming increasingly important for successful management of sharka disease. In this study, we have produced two super-sensitive and specific anti-PPV monoclonal antibodies (i.e., MAbs 13H4 and 4A11). Using these two MAbs, we have now developed a dot enzyme-linked immunosorbent assay (dot-ELISA) and a colloidal gold immunochromatographic strip (CGICS) assay. These two technologies can be used to quickly and reliably detect PPV. The results of these sensitivity assays confirmed that the dot-ELISA and CGICS assays could detect PPV infection in apricot tree leaf crude extracts diluted up to 1:5120 and 1:6400 (w/v), respectively. Further analyses using field-collected apricot tree leaf samples showed that the detection endpoint of the dot-ELISA was ~26 times above that obtained through RT-PCR, and the CGICS was as sensitive as RT-PCR. This present study is to broaden the knowledge about detection limits of dot-ELISA and CGICS for PPV monitoring. We consider that these newly developed dot-ELISA and CGICS are particularly useful for large scale PPV surveys in fields. Full article

A multicolor immunochromatographic assay platform based on quantum dot nanobeads (QBs) for the rapid and simultaneous detection of nitrofuran metabolites in different aquatic products is documented. These metabolites include 3-amino-2-oxazolidinone (AOZ), 1-aminohydantoin (AHD), semicarbazide (SEM), and 3-amino-5-morpholino-methyl-1,3-oxazolidinone (AMOZ). QBs with emission colors of red, yellow, green, and orange were employed and functionalized with the corresponding antibodies to each analyte to develop a multicolor channel. The visual detection limits (cutoff values) of our method for AOZ, AHD, SEM, and AMOZ reached up to 50 ng/mL, which were 2, 20, 20, and 20 times lower than those of traditional colloidal gold test strips, respectively. The test strip is capable of detection within 10 min in real samples while still achieving good stability and specificity. These results demonstrate that the developed multicolor immunochromatographic assay platform is a promising technique for multiplex, highly sensitive, and on-site detection of nitrofuran metabolites. Full article

Point-of-care testing methods for the rapid and sensitive screening of pathogenic bacteria are urgently needed because of the high number of outbreaks of microbial infections and foodborne diseases. In this study, we developed a highly sensitive and multiplex lateral flow assay (LFA) for the simultaneous detection of Pseudomonas aeruginosa and Salmonella typhimurium in complex samples by using wheat germ agglutinin (WGA)-modified magnetic quantum dots (Mag@QDs) as a universal detection nanoprobe. The Mag@QDs-WGA tag with a 200 nm Fe₃O₄ core and multiple QD-formed shell was introduced into the LFA biosensor for the universal capture of the two target bacteria and provided the dual amplification effect of fluorescence enhancement and magnetic enrichment for ultra-sensitivity detection. Meanwhile, two antibacterial antibodies were separately sprayed onto the two test lines of the LFA strip to ensure the specific identification of P. aeruginosa and S. typhimurium through one test. The proposed LFA exhibited excellent analytical performance, including high capture rate (>80%) to the target pathogens, low detection limit (<30 cells/mL), short testing time (<35 min), and good reproducibility (relative standard deviation < 10.4%). Given these merits, the Mag@QDs-WGA-based LFA has a great potential for the on-site and real-time diagnosis of bacterial samples. Full article

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. Full article

Tracking unreported allergens in commercial foods can avoid acute allergic reactions. A 2-step electrochemical immunosensor was developed for the analysis of the peanut allergen Ara h 1 in a 1-h assay (<15 min hands-on time). Bare screen-printed carbon electrodes (SPCE) were used as transducers and monoclonal capture and detection antibodies were applied in a sandwich-type immunoassay. The short assay time was achieved by previously combining the target analyte and the detection antibody. Core/shell CdSe@ZnS Quantum Dots were used as electroactive label for the detection of the immunological interaction by differential pulse anodic stripping voltammetry. A linear range between 25 and 1000 ng·mL⁻¹ (LOD = 3.5 ng·mL⁻¹), an adequate precision of the method (V_x0 ≈ 6%), and a sensitivity of 23.0 nA·mL·ng⁻¹·cm⁻² were achieved. The immunosensor was able to detect Ara h 1 in a spiked allergen-free product down to 0.05% (m/m) of peanut. Commercial organic farming cookies and cereal and protein bars were tested to track and quantify Ara h 1. The results were validated by comparison with an ELISA kit. Full article

A multiplex rapid detection system, based on a PCR-lateral flow biosensor (mPCR-LFB) was developed to identify Salmonella Typhi and Salmonella Paratyphi A from suspected carriers. The lower detection limit for S. Typhi and S. Paratyphi A was 0.16 and 0.08 ng DNA equivalent to 10 and 10² CFU/mL, respectively. Lateral flow biosensor was used for visual detection of mPCR amplicons (stgA, SPAint, ompC, internal amplification control) by labeling forward primers with fluorescein-isothiocyanate (FITC), Texas Red, dinitrophenol (DNP) and digoxigenin (DIG) and reverse primers with biotin. Binding of streptavidin-colloidal gold conjugate with the amplicons resulted in formation of a red color dots on the strip after 15–20 min of sample exposure. The nucleic acid lateral flow analysis of the mPCR-LFB was better in sensitivity and more rapid than the conventional agarose gel electrophoresis. Moreover, the mPCR-LFB showed 100% sensitivity and specificity when evaluated with stools spiked with 100 isolates of Salmonella genus and other bacteria. A prospective cohort study on stool samples of 1176 food handlers in outbreak areas (suspected carriers) resulted in 23 (2%) positive for S. Typhi. The developed assay has potential to be used for rapid detection of typhoid carriers in surveillance program. Full article

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. Full article

This work reviews the field of DNA biosensors based on electrochemical determination of nanoparticle labels. These labeling platforms contain the attachment of metal nanoparticles (NPs) or quantum dots (QDs) on the target DNA or on a biorecognition reporting probe. Following the development of DNA bioassay, the nanotags are oxidized to ions, which are determined by voltammetric methods, such as pulse voltammetry (PV) and stripping voltammetry (SV). The synergistic effects of NPs amplification (as each nanoprobe releases a large number of detectable ions) and the inherent sensitivity of voltammetric techniques (e.g., thanks to the preconcentration step of SV) leads to the construction of ultrasensitive, low cost, miniaturized, and integrated biodevices. This review focuses on accomplishments in DNA sensing using voltammetric determination of nanotags (such as gold and silver NPs, and Cd- and Pb-based QDs), includes published works on integrated three electrode biodevices and paper-based biosystems, and discusses strategies for multiplex DNA assays and signal enhancement procedures. Besides, this review mentions the electroactive NP synthesis procedures and their conjugation protocols with biomolecules that enable their function as labels in DNA electrochemical biosensors. Full article

Near-infrared fluorescence probes (NIFPs) have been widely used in immunoassay, bio-imaging and medical diagnosis. We review the basic principles of near-infrared fluorescence and near-infrared detection technology, and summarize structures, properties and characteristics of NIFPs (i.e., cyanines, xanthenes fluorescent dyes, phthalocyanines, porphyrin derivates, single-walled carbon nanotubes (SWCNTs), quantum dots and rare earth compounds). We next analyze applications of NIFPs in immunoassays, and prospect the application potential of lateral flow assay (LFA) in rapid detection of pathogens. At present, our team intends to establish a new platform that has highly sensitive NIFPs combined with portable and simple immunochromatographic test strips (ICTSs) for rapid detection of food-borne viruses. This will provide technical support for rapid detection on the port. Full article