Search Results (148)

Nanoporous gold nanoparticles (np-AuNPs) combine inertness, a nanoscale structure, and a porous framework with high surface area, conductivity, and biocompatibility, making them ideal for biosensing, catalysis, fuel cells, and drug delivery. Their open pore structure and low-coordinated atoms enhance biomolecule capture and mass transfer, while their tunable size, pore volume, and ease of surface modification make them promising biosensor transducers. However, synthesizing colloidal np-AuNPs in a simple way with controllable size and scalability remains challenging. The existing approaches mostly rely on specialized equipment, complex setups, and expert knowledge, while still facing challenges in terms of scalability. In this study, we present a simple, seedless, wet-chemical synthesis of colloidal np-AuNPs via the co-reduction of Au/Ag alloys followed by dealloying. By adjusting the Au:Ag ratio, we produced np-AuNPs sized ~120–530 nm, which were immobilized on electrodes for detecting lipopolysaccharide (LPS), a toxic component of Gram-negative bacterial membranes. The LPS biosensor exhibited excellent sensitivity towards detecting wild-type LPS, with a low limit of detection (LOD) of 0.1244 ng/L. This work demonstrates the effective synthesis and application of np-AuNPs in LPS biosensing. Full article

Co-infection of Orientia tsutsugamushi and influenza A virus complicates diagnosis and treatment in endemic regions because of overlapping clinical features and potential synergistic inflammation. We describe a 68-year-old woman from Hainan, China, who presented with five days of high fever (39.2 °C), nonproductive cough, eschar formation, lymphadenopathy, cytopenias, elevated liver enzymes, and raised inflammatory markers. On the day of admission, influenza A was confirmed by rapid antigen test and Orientia tsutsugamushi IgM/IgG was detected via colloidal-gold immunochromatography, prompting concurrent oseltamivir and doxycycline therapy. Quantitative PCR on day 2 measured an Orientia tsutsugamushi load of 2.85 × 10⁴ copies/mL (Cq 28.86), and targeted next-generation sequencing on day 3 revealed a high H1N1pdm09 viral burden (>1 × 10⁶ copies/mL) with low-level human herpesvirus 1 co-detection. Nested PCR and Sanger sequencing assigned Orientia tsutsugamushi to the Karp_A lineage and influenza A to clade 6B.1A.5a.2a. The patient defervesced by hospital day 2, laboratory indices normalized by day 3, and radiographic abnormalities resolved by day 6. This first documented Orientia tsutsugamushi–influenza A co-infection in China highlights the value of integrating rapid serology, qPCR quantification, nested PCR genotyping, and tNGS for early, precise dual-pathogen identification. Systematic multi-pathogen screening during overlapping transmission seasons is recommended to guide timely combination therapy and enhance epidemiological surveillance. Full article

Canine distemper, a fatal and highly transmissible disease caused by the canine distemper virus (CDV), poses a major threat to the companion animal industry. An urgent need exists for a rapid, specific, and simple method for the detection of this disease in order to improve its prevention and control. In this research, two monoclonal antibodies (mAbs), 1D3E9 and 1H9B7, were prepared, both of which specifically recognize the nucleoprotein (N protein) of CDV, and an immunochromatographic assay for CDV detection was subsequently developed using these mAbs. The results showed that both mAbs belong to the IgG1 subclass with kappa light chains. 1D3E9 was found to recognize the linear epitope ⁴¹⁰AGPKQSQITFLH⁴²¹, while 1H9B7 targeted the epitope ⁴⁵⁰HFNDERFPGH⁴⁵⁹. The test strips exhibited high specificity and good stability for up to two months when stored at 4, 25, and 37 °C. The assay exhibited a sensitivity of 10^2.39 TCID₅₀/0.1 mL. When compared with RT-PCR for detecting CDV in clinical samples, the concordance rate was 91.67%. Thus, this method shows great potential for facilitating rapid on-site detection of CDV and could be highly beneficial from the viewpoint of disease surveillance and control. Full article

In this study, a simple, mild, and eco-friendly cold plasma-solution interaction method is employed to rapidly prepare gold colloids. Through modification with multi-walled carbon nanotubes (MWCNTs), a non-enzymatic glucose-sensing electrode material is successfully fabricated. The prepared electrode material is characterized via X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), and transmission electron microscopy (TEM). The results show that compared with the chemically reduced AuNPs-C-MWCNTs, the plasma-prepared AuNPs-P-MWCNTs exhibits enhanced glucose catalytic performance with a higher sensitivity of 73 μA·mM⁻¹·cm⁻² (approximately 3.2 times that of AuNPs-C-MWCNTs), lower response time of 2.1 s, and ultra-low detection limit of 0.21 μM. It also demonstrates excellent selectivity, reproducibility (RSD = 4.37%), repeatability (RSD = 3.67%), and operational stability (RSD = 4.51%). This improvement can be attributed to the smaller particle size and better dispersion of plasma-derived AuNPs on the surface of MWCNTs. Furthermore, the AuNPs-P-MWCNTs surface is enriched with oxygen-containing functional groups, which is conducive to the enhancement of the hydrophilicity of the electrode surface. These synergistic effects facilitate the AuNPs-catalyzed glucose oxidation reaction, ultimately leading to superior glucose catalytic performance. Full article

This study aimed to develop monoclonal antibodies (mAbs) against the gD protein of FHV-1 for rapid and specific virus detection. The gD protein, a highly conserved part of the FHV-1 envelope, is crucial for viral entry into host cells, making it an ideal detection target. We immunized BALB/c mice with an mRNA vaccine encoding the gD gene, achieving a serum antibody titer of 1:140,000 after three immunizations. The mice were then boosted with recombinant gD protein. Through cell fusion and multiple subcloning rounds, we obtained five hybridoma cell lines (D7, E4, E9, E10, and E19) that stably secrete anti-gD protein mAbs. Characterization by indirect immunofluorescence and Western blot showed that mAbs D7 and E4 have high specificity and strong binding activity against FHV-1, detectable at 2 μg/mL. These mAbs provide specific tools for FHV-1 detection and a basis for developing rapid diagnostic methods using ELISA, colloidal gold, and other technologies. Full article

Background/Objectives: The COVID-19 pandemic has highlighted the urgent need for rapid, accurate, and accessible diagnostic testing to effectively manage and contain the spread of SARS-CoV-2. RT-PCR is widely recognized as the gold standard for SARS-CoV-2 detection due to its high sensitivity and specificity. However, RT-PCR testing requires specialized laboratory equipment, highly trained personnel, and extended processing times, which limits its feasibility for large-scale screening and point-of-care applications. This study aims to systematically evaluate the diagnostic performance of RT-PCR and a colloidal gold immunochromatographic assay (GICA). Methods: By comparing these two methods, we seek to determine a GICA’s effectiveness as a complementary or alternative diagnostic tool, particularly in resource-limited settings and scenarios requiring rapid, large-scale testing. We assessed the following key clinical parameters: sensitivity, specificity, NPV, PPV, and accuracy. Additionally, we investigated the correlation between GICA signal intensity and RT-PCR Ct values using regression analysis, receiver operating characteristic curve analysis, and the calculated area under the curve. Results: Our findings indicate that while RT-PCR exhibits superior sensitivity, GICA results demonstrate a strong correlation with RT-PCR results and provide a rapid, cost-effective alternative for SARS-CoV-2 detection. Unlike RT-PCR, which requires extensive resources and prolonged turnaround times, a GICA delivers results within 20 min, making it a viable option for decentralized testing and real-time public health interventions. Conclusions: These results suggest that a GICA can serve as a complementary diagnostic tool alongside RT-PCR, particularly in resource-limited settings and high-throughput screening scenarios. By integrating GICAs into broader testing strategies, healthcare systems can enhance early detection efforts, improve accessibility to diagnostics, and strengthen pandemic response measures. Full article

Toxoplasma gondii is a globally significant zoonotic pathogen responsible for severe parasitic diseases in humans and animals. This study aimed to design, develop, and evaluate a novel immunochromatographic test (ICT) using a recombinant MIC2-MIC3 fusion protein (rMIC2-MIC3) for detecting specific antibodies against T. gondii. The ICT demonstrated exceptional sensitivity, capable of detecting T. gondii-specific antibodies in sera diluted up to 1:8. Specificity evaluation confirmed no cross-reactivity with antibodies against other parasites, such as Neospora caninum, Cryptosporidium suis, Eimeria tenella, and Sarcocystis tenella. Stability tests revealed the test strips maintained full functionality after 12 weeks of storage at 24 °C. The coincidence rate of the colloidal gold test strips prepared in this study with a commercial ELISA kit was 94.59%. Comparisons with advanced serodiagnostic tools, such as chimeric antigen-based ELISAs and recombinant protein diagnostics, further highlighted its robustness and applicability. These findings underscore the potential of the rMIC2-MIC3-based ICT as a reliable, economical, and accessible diagnostic tool for toxoplasmosis in veterinary and human medicine. Full article

The optical detection of thioacetamide was investigated using a metalated porphyrin, Mn(III)-5,10,15,20-tetrakis-(3,4-dimethoxyphenyl)-21H,23H-porphyrin chloride (Mn-3,4-diMeOPP), a gold colloid solution (AuNPs), and a complex formed between them (Mn-3,4-diMeOPP–AuNPs) in order to select the most sensitive material and to achieve complementarity between methods. Mn-3,4-diMeOPP, AuNPs, and their complex were synthesized and characterized by means of UV–Vis, FT-IR spectrometry, and AFM investigations. It could be concluded that Mn-3,4-diMeOPP could detect/quantify thioacetamide (TAA) in the range 3.13 × 10⁻⁸ M–7.67 × 10⁻⁷ M in a linear fashion, with a 99.85% confidence coefficient. The gold colloidal particles alone could detect TAA in an extremely narrow concentration domain of 2–9.8 × 10⁻⁷ M, slightly complementary with that of Mn-3,4-diMeOPP. The complex between Mn-3,4-diMeOPP and gold colloid proved to be able to quantify TAA in the trace domain with concentrations of 1.99 × 10⁻⁸ M–1.76 × 10⁻⁷ M in a polynomial fashion, with the method being more difficult. A potential mechanism for TAA detection based on Mn-3,4-diMeOPP is discussed based on computational modeling. The distorted porphyrin conformation and its electronic configuration favor the generation of a grid of electrostatic interactions between porphyrin and TAA. Full article

E. coli O157:H7 contamination in food and the environment poses a serious threat to human health. Rapid and sensitive identification of foodborne pathogens remains challenging. Here, we prepared tungsten disulfide (WS₂)–Au nanocomposites coupled with the Raman signal molecule 5,5′-dithio-bis-(2-nitrobenzoic acid) (DTNB) and antibodies to replace the conventional colloidal gold nanoparticles and applied SERS-active nanotags in the SERS-ICA method for highly sensitive detection of E. coli O157:H7. The large surface area and numerous effective SERS hotspots of WS₂-Au nanotags provide superior SERS signals. Under optimized conditions, this ICA achieves the quantitative detection of E. coli O157:H7 in a broad linear range of 8 × 10²–8 × 10⁷ CFU/mL and at a low detection limit of 175 CFU/mL. In addition, the test strip indicates high specificity for E. coli O157:H7 identification, favorable reproducibility, and shows good accuracy in the detection of actual food samples, such as milk and pork. The proposed assay can be used for rapid qualitative and quantitative detection of E. coli O157:H7 and has great potential for field application. Full article

The clinical characteristics of multidrug-resistant tuberculosis (MDR-TB) patients with concurrent nontuberculous mycobacterial (NTM) infection present significant challenges to treatment. This study investigated the clinical characteristics of MDR-TB patients with concurrent NTM infection during treatment. A retrospective cohort study was conducted to collect the clinical data of MDR-TB patients who initiated treatment between January 2020 and December 2022. A total of 389 patients were analyzed, among which 111 patients who were lost to follow-up and 56 patients who missed etiological examination of tuberculosis during the visit period were excluded. A total of 222 patients with complete data were included in this study. The species identification method primarily employed molecular biology techniques, specifically the DNA microarray method and/or MPB64 antigen detection using the colloidal gold method. Patients whose sputum or bronchoalveolar lavage fluid cultures were positive and who were identified at least once as having NTM or as MPB64 negative were included in this study. Imaging data, comorbidities, pre-treatment infection, and nutritional indicators were analyzed during treatment. Among the 222 MDR-TB patients, no concurrent NTM cases were identified at the beginning of treatment. However, 19 cases (8.56%) were presumed to be NTM-positive during treatment, which appeared during anti-tuberculosis treatment from 2 to 12 months, averaging 6 (3, 12) months. Thirteen patients were only tested for MPB64, with five having two negative MPB64 tests. The symptoms of NTM-positive patients varied, and imaging findings were similar to those of MDR-TB but did not worsen. The emergence of presumed NTM-positive cases (8.56%) among MDR-TB patients during treatment highlights the need for monitoring, as symptoms and imaging findings may mimic MDR-TB without worsening. Early and repeated testing, including methods beyond MPB64, may be useful for more accurate diagnosis and tailored management. Full article

Actinobacillus pleuropneumoniae (APP) is a bacterial pathogen causing porcine pleuropneumonia, causing great economic loss to the global pig industry. Although natural apxIV contributes to the prevention and control of porcine pleuropneumonia, its isolation poses a great challenge, and recombinant soluble apxIV proteins tend to carry large molecular weight tags. The traditional serologic methods tend not to accurately detect the apxIV-partially deleted vaccine (GDV). In this study, we screened the soluble protein apxIVA N2 (756 bp) from six apxIV-truncated proteins and applied it to the enzyme-linked immunosorbent assay (ELISA) and colloidal gold immunochromatographic strip for detecting the samples vaccinated with APP GDV. The results indicate that N2 was close to the natural apxIV protein in terms of structure and function as it only contained a single His (0.86 kDa) tag and a single S (2 kDa) tag. Among the six candidate proteins, N2 exhibited the best performance in distinguishing APP-infected samples from those vaccinated with the APP GDV. Both ELISA and colloidal gold immunochromatographic strips based on this protein exhibited an excellent performance in detecting and distinguishing wild-strain-infected samples from those vaccinated with the subunit vaccine or the GDV. In addition, three monoclonal antibodies against different antigenic epitopes were identified using these truncated proteins. Our studies are of great significance for further research on APP, the differential diagnosis of wild strains and vaccine strains, and pig control breeding, exhibiting a broad application prospect in the on-site diagnosis of APP, particularly in remote areas lacking detection instruments and professionals. Full article

The quantitative analysis of nucleic acid markers is extensively utilized in cancer detection. However, it faces significant challenges, such as the need for specialized detection devices and the inherent complexity of testing procedures. To address these issues, this study proposes a simplified, rapid, and user-friendly platform for cancer nucleic acid marker detection. We firstly designed a polydimethylsiloxane (PDMS) device for the isothermal amplification reaction of nucleic acid biomarkers based on reverse-transcription recombinase-aided amplification (RT-RAA) technology. Specifically, three potential cancer nucleic acid biomarkers, carcinoembryonic antigen (CEA), prostate-specific antigen (PSA), and prostate cancer antigen 3 (PCA3) were amplified from human serum or urine samples in the PDMS device at body temperature. The reaction chamber was directly integrated with nucleic acid test strips labeled with colloidal gold nanoparticles, allowing for the visual observation of the detection results for the amplification products. The optimal reaction conditions, such as pH, reaction time, antibody, and streptavidin concentration, were defined after a series of optimization studies. The findings demonstrated that the optimal RT-RAA reaction time was 20 min, the primary antibodies were labeled with colloidal gold to the greatest extent at pH 8.5, and the optimal concentrations of secondary antibody and streptavidin were 1.0 mg/mL and 0.5 mg/mL, respectively. Furthermore, this novel detection approach could not only exhibit excellent sensitivity and specificity but also show high accuracy for the analysis of nucleic acid biomarkers in both clinical serum and urine samples. Therefore, the simplified and more convenient operation platform provides a new insight for the semi-quantitative analysis of cancer nucleic acid biomarkers and the rapid screening of early cancer, thereby offering a promising alternative to oncological point-of-care testing (POCT) diagnostics. Full article

Among algae that synthesize paralytic shellfish toxins (PSTs), Alexandrium tamarense is a widely distributed and highly dangerous species with significant impacts on the marine environment and human health. Therefore, establishing fast and reliable monitoring technology for Alexandrium tamarense is crucial. Developing effective detection and early warning systems for toxic red tides is of paramount importance. Conventional detection methods, such as microscopy and molecular biology, are complex and time-consuming, requiring specialized personnel and equipment, which makes them unsuitable for on-site rapid testing. In this study, we successfully developed polyclonal and monoclonal antibodies targeting Alexandrium tamarense using colloidal gold immunochromatography technology. Based on these antibodies, we created colloidal gold test strips capable of detecting Alexandrium tamarense in water samples. These test strips enable rapid detection of the target algae in aquatic environments and semi-quantitative estimation of algal concentrations using a colorimetric card. They can quickly determine whether the concentration of red tide algae has reached a critical level, allowing for timely preventive measures. This innovation holds significant practical value and broad application potential. Full article

Background: During the COVID-19 pandemic, we continuously monitored the epidemiology of influenza virus among pediatric patients from January 2021 to December 2023 in Hainan Island, China. Methods: In this study, we collected 54,974 nasopharyngeal swab samples for influenza A Virus (IAV) testing and 53,151 samples for influenza B Virus (IBV) testing from pediatric outpatients. Additionally, we also collected 19,687 nasopharyngeal swab samples from pediatric inpatients for IAV and IBV testing. Outpatient samples were screened for influenza viruses (IVs) infection by the colloidal gold method. Targeted Next-Generation Sequencing (tNGS) was used to detect influenza virus infections in inpatients. Influenza virus types were identified by analyzing the HA/NA partial regions. Results: The findings revealed a significant decrease in the infection rate of IBV over the specified period, while the infection rate of IAV exhibited a rising trend. Additionally, B/Victoria lineage was the dominant epidemic strain in 2021, while the epidemic strains in 2022 and 2023 underwent a dynamic transformation from A/H3N2 to A/H1N1. Phylogenetic analysis revealed close relationships among the circulating strains. Nonetheless, because the sample size is limited, additional research is required. Conclusions: Our findings suggest that the predominant types of influenza viruses in the pediatric population are undergoing dynamic changes, influenced by the implementation and relaxation of non-pharmaceutical intervention measures. These findings highlight the need for adaptive influenza vaccination and containment strategies, particularly in tropical regions like Hainan, where climate and public health policies significantly impact viral transmission patterns. The insights gained from this study could inform more effective public health strategies in similar regions to mitigate the impact of influenza outbreaks in the future. Full article

Pesticide residues still pose a risk to human health. With the rapid development of rapid testing technology, the levels of different types of pesticide residues in agricultural products can be identified in a shorter period; thus, the safety of food can be guaranteed. However, the effectiveness of commercially available testing products has yet to be evaluated. In this study, colloidal gold immunochromatographic qualitative testing products manufactured by 34 companies were tested for their assay performance on Emamectin Benzoate, Isocarbophos, and fipronil with standardized cowpea samples. The results indicated that most of the evaluated products were identified as having ‘passed’. Most pesticide residue rapid test immunoassay products can be considered ideal means for testing certain pesticide residues. However, further evaluation of pesticide residue rapid test immunoassay products is needed, as detection technologies are still developing. Full article