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The escalating issue of drug abuse poses a significant threat to public health and societal stability worldwide. An on-site drug detection platform is vital for combating drug abuse and trafficking, as it eliminates the need for additional tools, extensive processes, or specialized training. Therefore, it is imperative to develop a fast, sensitive, non-invasive, and reliable multiplex drug testing platform. In this study, we have presented a silica core@dual quantum dot-shell nanocomposite (SI/DQD)-based fluorescent lateral flow immunoassay (LFIA) platform for the highly sensitive and simultaneous point-of-care (POC) detection of methamphetamine (MET) and tramadol (TR). A 3D-printed attachment was designed to integrate optical and electrical components, facilitating the miniaturization of the instrument and reducing both cost and complexity. The device's advanced hardware and effective fluorescence extraction algorithm with waveform reconstruction enable swift, automatic noise reduction and data analysis. SI/DQD nanocomposites were utilized as fluorescent nanotags in the LFIA strips due to their outstanding luminous efficiency and robustness. This LFIA platform achieves impressive detection limits (LODs) of 0.11 ng mL -1 for MET and 0.017 ng mL -1 for TR. The method has also successfully detected MET and TR in complex biological samples, demonstrating its practical application capabilities. The proposed fluorescent LFIA platform, based on SI/DQD technology, holds significant promise for the swift and accurate POC detection of these substances. Its affordability, compact size, and excellent analytical performance make it suitable for on-site drug testing, including at borders and roadside checks, and open up new possibilities for the design and implementation of drug testing methods., Competing Interests: Declaration of competing interest No conflict of interest exists in the submission of this manuscript, and the manuscript is approved by all authors for publication., (Copyright © 2024 Elsevier B.V. All rights reserved.)

A point-of-care testing biosensor that supports direct, sensitive, and simultaneous identification of bacteria and virus is still lacking. In this study, an ultrasensitive immunochromatography assay (ICA) with colorimetric/fluorescence dual-signal output was proposed for flexible and accurate detection of respiratory virus and bacteria in complex samples. Colorimetric AuNPs of 16 nm and two layers of quantum dots (QDs) were coated onto the surface of monolayer graphene oxide (GO) layer by layer to form a multilayered dual-signal nanofilm. This material not only can generate strong colorimetric and fluorescence signals for ICA analysis but also can provide larger surface area, better stability, and superior dispersibility than conventional spherical nanomaterials. Two test lines were built onto the ICA strip to simultaneously detect common respiratory virus influenza A and respiratory bacteria Streptococcus pneumoniae. The dual-signal mode of assay greatly broadened the applied range of ICA method, in which the colorimetric mode allows for quick determination of virus/bacteria and the fluorescence mode ensures the highly sensitive and quantitative detection of target pathogens with detection limits down to 891 copies/mL and 17 cells/mL, respectively. The proposed dual-mode ICA can also be applied directly for real biological and environment samples, which suggests its great potential for field application., Competing Interests: Declaration of Competing Interest 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., (Copyright © 2023 Elsevier B.V. All rights reserved.)

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.

A rapid and accurate method for detection of virus (SARS-CoV-2)-specific antibodies is important to contain the 2019 coronavirus disease (COVID-19) outbreak, which is still urgently needed. Here, we develop a colorimetric-fluorescent dual-mode lateral flow immunoassay (LFIA) biosensor for rapid, sensitive, and simultaneous detection of SARS-CoV-2-specific IgM and IgG in human serum using spike (S) protein-conjugated SiO 2 @Au@QD nanobeads (NBs) as labels. The assay only needs 1 μL of the serum sample, can be completed within 15 min, and is 100 times more sensitive than the colloidal gold-based LFIA. Two detection modes of our biosensor are available: the colorimetric mode for rapid screening of the patients with suspected SARS-CoV-2 infection without any special instrument and the fluorescent mode for sensitive and quantitative analyses to determine the concentrations of specific IgM/IgG in human serum and detect the infection early and precisely. We validated the proposed method using 16 positive serum samples from patients with COVID-19 and 41 negative samples from patients with other viral respiratory infections. The results demonstrated that combined detection of virus-specific IgM and IgG via SiO 2 @Au@QD LFIA can identify 100% of patients with SARS-CoV-2 infection with 100% specificity.

Novel diagnostic tools are a major challenge for brucellosis research, especially in developing countries. Herein, we established a handheld quantum dot (QD) immunochromatographic device for the fast detection of brucellosis antibodies in the field. Total bacterial protein extracted from Brucella 104M served as labelling and coating antigen. QD labelling and immunochromatography methods were used to optimise reaction conditions, labelling conditions, reaction temperature and storage temperature. QD test strips were employed to test brucellosis serum to determine their sensitivity, specificity and stability. Test strips were compared with Rose Bengal test, standard agglutination test and colloidal gold immunochromatographic assay. Labelled Brucella total protein displayed good specificity and no cross-reactivity. The concentration of labelled total bacterial protein was 3.9 mg/ml, the coating concentration was 2.0 mg/ ml, and the serum titre with the lowest detection sensitivity was 1:25. The optimal reaction temperature for the test strip was 25-30°C. The test strip was stable after storage at room temperature and the repeatability was high, with a coefficient of variation of 4.0%. After testing 199 serum samples, the sensitivity of the QD test strip was 98.53%, the specificity was 93.57%, and the coincidence rate with the standard agglutination test was 96.98%. The developed QD immunochromatographic method can be used for rapid detection and preliminary screening of brucellosis in the field., Competing Interests: The authors have declared that no competing interests exist.

Lateral flow immunoassay (LFA) strips are extensively used for rapid tests of various biochemical molecules, but these strips still have some limitations in bacterial detection due to their low sensitivity and poor stability in complex samples. In this study, we reported a highly sensitive and quantitative fluorescent LFA strip for bacterial detection by using novel magnetic-core@dual quantum dot (QD)-shell nanoparticles (Fe3O4@DQDs) as multifunctional fluorescent labels. The Fe3O4@DQDs were prepared through a polyethyleneimine (PEI)-mediated layer-by-layer (LBL) assembly method, and they possess monodispersity, high magnetic responsiveness, good stability, and superior fluorescence properties. Based on these merits, the Fe3O4@DQDs were used to capture and enrich bacteria from complex samples and then used as advanced fluorescent labels of LFA strips for the quantitative detection of bacteria. Under optimal conditions, the assay ultra-sensitively detected Streptococcus pneumoniae with a low limit of detection of 8 cells per mL and a wide dynamic linear range of 10 cells per mL to 107 cells per mL. Systematic comparison revealed that the fluorescence detection limit of the Fe3O4@DQD-based strip was 55 and 1000 times higher than those of Fe3O4-core@QD-shell nanocomposite (Fe3O4-QD)-based and conventional QD microsphere-based strips, respectively. The proposed method also exhibited high specificity and selectivity for biological samples (human whole blood and sputum) and is thus a promising tool for real bacterial sample testing.

Protein toxins, such as botulinum neurotoxin type A (BoNT/A) and staphylococcal enterotoxin B (SEB), easily pollute food and water and are ultra-toxic to humans and animals, thus requiring a sensitive on-site detection method. In this study, we reported a novel lateral flow assay (LFA) strip on the basis of magnetic quantum dot nanoparticles (MagQD NPs) for sensitive and multiplex protein toxin detection in food samples. A new type of MagQD NP was prepared by fixing the dense carboxylated QDs on the surface of polyethyleneimine-modified Fe 3 O 4 magnetic NPs (MNPs) and applied in LFA with the following functions: capture and enrich target toxins from sample solutions and serve as advanced fluorescent labels for the quantitative determination of targets on the strip. Through this strategy, the assay realized quantified BoNT/A and SEB detection in 30 min with the limits of detection of 2.52 and 2.86 pg/mL, respectively. The selectivity and the ability of quantitative analysis of the method were validated in real food samples, including milk and juice. This MagQD-LFA biosensor showed considerable potential as a point-of-care testing tool for the sensitive detection of trace toxins., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This work describes an anti-ovalbumin antibody-based lateral flow immunoassay (LFI) for T-2 toxin. The antibody uses a coating antigen as a bifunctional element for universality and introduces preincubation to improve the detection limits of the method. T-2 toxin and ovalbumin-modified T-2 toxin competitively binds on the anti-T-2 toxin monoclonal antibody modified on CdSe/ZnS quantum dot beads during preincubation. The modified T-2 toxin acts as a bifunctional element that forms immuno complexes during preincubation and combines with anti-ovalbumin antibody coated in the test line through the ovalbumin terminal. Fluorescence is detected at 610 nm on the test zone following photoexcitation at 365 nm. It has a reverse dose-effect relationship with the amount of T-2 toxin. The calibration plot is linear in the 20-110 fg mL -1  T-2 toxin concentration range, and the limit of detection (LOD) is 10 fg mL -1 , which is lower by 8-fold than that of the traditional LFI system (LOD 80 fg mL -1 ) and one order of magnitude than those of LFIs with labels of colloidal gold nanoparticles (LOD 150 fg mL -1 ) or fluorophores (LOD 190 ng mL -1 ). Universality was verified through aflatoxin B 1 detection using the established ovalbumin antibody-based LFI system (LOD 10 fg mL -1 ). The performance of the method was compared with that of established systems and a commercial ELISA kit (LOD 360 fg mL -1 ). Graphical abstractSchematic representation of ovalbumin antibody-based immunochromatographic lateral flow assay for T-2 toxin. Preincubation is introduced for high sensitivity. T-2- anti-ovalbumin acts as a bi-functional element for universality. CdSe/ZnS quantum dot beads act as label. Fluorometric signal is detected at 610 nm.

MiRNAs play important regulatory roles in numerous biological processes and serve as significant biomarkers for the development and prognosis of several diseases. Their unique characteristics, such as short size, high sequence homology among family members, low abundance, and easy degradability, have hindered their specific and highly sensitive detection. Herein, a duplex-specific nuclease (DSN)-assisted target recycling signal amplification-based fluorescent lateral flow assay was demonstrated for the point-of-care detection of cancer-related miRNA-21. In this assay, digoxin/biotin-labeled DNA probes were selectively cleaved by the DSN enzyme in the rounds of hybridization with the miRNA-21 target and cleavage cycle. Subsequently, the resulting mixture, containing the miRNA-21 target and intact and cleaved DNA probes, was loaded onto the lateral flow strip with digoxin antibody-conjugated quantum dot nanobeads and the streptavidin-coated test line. The increase in the proportion of cleaved DNA probes can induce a weakened response signal, which is directly associated with the amount of the miRNA target. Thus, highly sensitive quantification of miRNA-21 was achieved at a low limit of detection of 0.16 pM within 2 h of assay time. Assay specificity toward miRNA-21 was validated by testing several other miRNAs, including let-7b, let-7d, miRNA-141, and miRNA-200a. Moreover, the assay can quantify miRNA-21 spiked in human serum samples with acceptable recovery values, thus indicating its considerable clinical feasibility. [ABSTRACT FROM AUTHOR]

There is a strong need for the development of a sensitive biosensor for real-time monitoring mycotoxins contamination in real food samples. Here, we reported a multiplexed immunochromatographic assay (ICA) using functionalized magnetic-quantum dot (QD) as capture/detection difunctional immunoprobes that can simultaneously and sensitively detect aflatoxin B1 (AFB1), ochratoxin A (OTA), and fumonisin B1 (FB1) in food. The antibodies-modified multilayer magnetic-QD nanobeads (MagQBD) allowed effective capture of three target mycotoxins from complex solution and simultaneously detect them on one ICA strip through fluorescence signal reading. The detection sensitivity of MagQBD-ICA was markedly improved due to the signal amplification of the multilayer QD shell and target enrichment. With capture/detection strategy, multiplexed detection of AFB1, OTA, and FB1 was conducted, resulting in low detection limits of 0.42, 11.48, and 4.21 pg mL–1, respectively. The excellent accuracy, specificity, and stability of the method were further confirmed by testing actual samples including rice, peanut, corn and juices. Results suggested the great potential of MagQBD-ICA for practical detection of mycotoxins-contaminated food. [Display omitted] • Magnetic-fluorescence ICA biosensor for multiplex detection of mycotoxins was first proposed. • Multilayer magnetic-QD nanobead was employed as capture/detection difunctional immunoprobes. • Accurate and sensitive identification of multiple mycotoxins in real foods was achieved. • The detection limits for three target mycotoxins reached pg/mL level. [ABSTRACT FROM AUTHOR]

[Display omitted] • Two-dimensional GO-QD nanoflakes were fabricated as flexible nanoprobes for bacteria. • GO-based nanoprobe with fluorescence-enhanced ability was firstly introduced into ICA strip. • The developed biosensor detected S. pneumoniae and S. aureus with LOD values of 13 and 20 cells/mL, respectively. • This is the first work to achieve two respiratory bacteria simultaneous detection via ICA. Respiratory pathogens including a variety of bacteria and viruses are highly infectious to human, cause similar clinical symptoms, and pose serious threats to public health globally. Early and accurate identification of respiratory bacteria from other pathogens is crucial for guiding the rational use of antibiotics and controlling the epidemic spread. Here, we developed a multiplexed fluorescent immunochromatographic assay (ICA) for ultrasensitive qualification of respiratory bacteria in biological samples with high accuracy. A novel two-dimensional graphene oxide (GO)-based quantum dot (QD) nanoflake was designed as film-like immunoprobe to efficiently and closely stick onto target bacteria surface, which not only provide superior fluorescence signal but also ensure the liquidity of the captured bacteria for ICA detection. By combining the GO-QD nanoflakes and ICA strip, the proposed method enables the simultaneous detection of two common respiratory bacteria, namely Streptococcus pneumoniae (S. pneumoniae) and Staphylococcus aureus (S. aureus) in biological samples with low detection limit (13, 20 cells/mL), fast detection speed (20 min), good reproducibility (RSD < 4.2 %), and high specificity. The excellent performance of our proposed GO-QD-ICA suggests its great potential for rapid and ultrasensitive diagnosis of real respiratory tract samples. [ABSTRACT FROM AUTHOR]

• Ultrasensitive fluorescent ICA for rapid screening of respiratory viruses were established. • Multilayer silica-QD nanobead with excellent stability and superior luminescence was fabricated. • The proposed method detected SARS-CoV-2 NP and FluA H1N1 with LOD values of 5 pg/mL and 50 pfu/mL, respectively. • This is the first work to achieve SARS-CoV-2 and FluA antigens simultaneous detection via ICA. Rapid and accurate diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and influenza A virus (FluA) antigens in the early stages of virus infection is the key to control the epidemic spread. Here, we developed a two-channel fluorescent immunochromatographic assay (ICA) for ultrasensitive and simultaneous qualification of the two viruses in biological samples. A high-performance quantum dot nanobead (QB) was fabricated by adsorption of multilayers of dense quantum dots (QDs) onto the SiO 2 surface and used as the highly luminescent label of the ICA system to ensure the high-sensitivity and stability of the assay. The combination of monodispersed SiO 2 core (∼180 nm) and numerous carboxylated QDs formed a hierarchical shell, which ensured that the QBs possessed excellent stability, superior fluorescence signal, and convenient surface functionalization. The developed ICA biosensor achieved simultaneous detection of SARS-CoV-2 and FluA in one test within 15 min, with detection limits reaching 5 pg/mL for SARS-CoV-2 antigen and 50 pfu/mL for FluA H1N1. Moreover, our method showed high accuracy and specificity in throat swab samples with two orders of magnitude improvement in sensitivity compared with traditional AuNP-based ICA method. Hence, the proposed method is a promising and convenient tool for detection of respiratory viruses. [ABSTRACT FROM AUTHOR]

• Magnetic-quantum dot nanobeads can serve as enrichment substrate and fluorescent probe for virus detection using lateral flow assay. • Influenza A virus (IAV) virions were separated and enriched from clinical specimens. • Ultrasensitive detection of IAV virions with a LOD down to 22 pfu mL–1 within 35 minutes. • Good specificity between influenza B virus and two adenovirus strains. Influenza A virus (IAV) possesses a high infectivity and pathogenicity, and can lead to severe respiratory infection with similar symptoms caused by some other common respiratory viruses. Lateral flow assay (LFA) has been widely deployed in remote settings as a rapid and reliable approach for point-of-care detection of infectious pathogens. However, it still remains challenging to detect IAV virions using LFA from clinical samples such as nasopharyngeal or throat swabs, because their various components and high viscosity can decrease flow velocity and lead to the nonspecific adsorption of nanoparticle labels on the sensing membrane. Herein, we demonstrated a magnetic quantum dot nanobeads (MQBs) based LFA for magnetic enrichment and fluorescent detection of IAV virions in clinical specimens. In this study, MQBs were synthesized and then conjugated with IAV-specific antibody to efficiently enrich IAV virions from complex biological matrix, but also serve as highly bright fluorescent probes in lateral flow strips. This assay can achieve quantitative detection of IAV virions with a low limit of detection down to 22 pfu mL–1 within 35 minutes, and show good specificity between influenza B virus and two adenovirus strains. Furthermore, the presented platform was able to directly detect IAV virions spiked in nasopharyngeal swab dilution, indicating its stability and feasibility in clinical applications. Thus, this point-of-care detection platform holds great promise as a broadly applicable approach for the rapid diagnosis of influenza A. [ABSTRACT FROM AUTHOR]