Plasmonic Approach to Fluorescence Enhancement of Mesoporous Silica-Coated Gold Nanorods for Highly Sensitive Influenza A Virus Detection Using Lateral Flow Immunosensor.

Rapid diagnostic tests based on the lateral flow immunoassay (LFI) enable early identification of viral infection, owing to simple interpretation, short turnaround time, and timely isolation of patients to minimize viral transmission among communities. However, the LFI system requires improvement in the detection sensitivity to match the accuracy of nucleic acid amplification tests. Fluorescence-based LFIs are more sensitive and specific than absorption-based LFIs, but their performance is significantly affected by fundamental issues related to the quantum yield and photobleaching of fluorophores. Metal-enhanced fluorescence (MEF), which is a plasmonic effect in the vicinity of metallic nanoparticles, can be an effective strategy to improve the detection sensitivity of fluorescence-based LFIs. The key factors for obtaining a strong plasmonic effect include the distance and spectral overlap of the metal and fluorophore in the MEF system. In this study, MEF probes were designed based on core-shell nanostructures employing a gold nanorod core, mesoporous silica shell, and cyanine 5 fluorophore. To optimize the efficiency of MEF probes incorporated on the LFI platform (MEF-LFI), we experimentally and theoretically investigated the distance dependence of plasmonic coupling between cyanine 5 and gold nanorods by adjusting the shell thickness, resulting in significant fluorescence enhancement. The proposed MEF-LFI enabled highly sensitive detection of influenza A virus (IAV) nucleocapsid protein with a detection limit of 0.52 pg mL-1 within 20 min and showed high specificity and accuracy for determining IAV clinical samples. Overall, our findings demonstrate the potential of this method as an effective tool for molecular diagnosis under emergency conditions.

Absorption-Modulated SiO2@Au Core-Satellite Nanoparticles for Highly Sensitive Detection of SARS-CoV-2 Nucleocapsid Protein in Lateral Flow Immunosensors.

The worldwide spread of coronavirus disease 2019 (COVID-19) highlights the need for rapid, simple, and accurate tests to detect various variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The antigen test, based on the lateral flow immunoassay (LFI), is a suitable "first line of defense" test that enables early identification and timely isolation of patients to minimize viral transmission among communities. However, it is generally less accurate than nucleic acid testing, and its sensitivity needs improvement. Here, a novel rapid detection method is designed to sensitively detect SARS-CoV-2 using isolated gold nanoparticle (AuNP)-assembled SiO2 core-satellite nanoparticles (SiO2@Au CSNPs). Well-grown AuNP satellites in the synthesis of SiO2@Au CSNPs significantly enhanced their light absorption, increased the detection sensitivity, and lowered the detection limit by 2 orders of magnitude relative to conventional gold colloids. The proposed system enabled highly sensitive detection of the SARS-CoV-2 nucleocapsid protein with a detection limit of 0.24 pg mL-1 within 20 min. This is the first study to develop a highly sensitive antigen test using the absorption-modulated SiO2@Au CSNPs. Our findings demonstrate the capacity of this platform to serve as an effective sensing strategy for managing pandemic conditions and preventing the spread of viral infections.

Direct Use of a Saliva-Collected Cotton Swab in Lateral Flow Immunoassay for the Detection of Cotinine.

The detection of salivary cotinine is useful for convenient smoking tests in spite of the high background effect of saliva. For precise results, the conventional salivary cotinine analysis for smoking detection requires complex pretreatment processes. Hence, in this study, we developed a modified paper-based lateral flow immunoassay (LFIA), termed "gap-LFIA", for the direct application of saliva collected using cotton swabs for on-site detection. The gap-LFIA was constructed by modifying a conventional LFIA sensor, where the sample pad was divided to have a 3 mm gap. A saliva-collected cotton swab was inserted into the gap, and then, a buffer solution was added to the outer sample pad to dilute the saliva automatically. The gap-LFIA reduced the interference in salivary samples and showed improved signals, allowing for using the whole saliva directly without additional steps. Further, the deviation of results using a strip was less than that when the saliva was not diluted in a conventional cotinine kit, and it helped to distinguish between smokers and non-smokers more clearly in 15 min. This method of automatic dilution may apply to various clinical samples, including blood and serum, for direct application in future detections.

Enhancement of the catalytic activity of ferulic acid decarboxylase from Enterobacter sp. Px6-4 through random and site-directed mutagenesis.

The enzyme ferulic acid decarboxylase (FADase) from Enterobacter sp. Px6-4 catalyzes the decarboxylation reaction of lignin monomers and phenolic compounds such as p-coumaric acid, caffeic acid, and ferulic acid into their corresponding 4-vinyl derivatives, that is, 4-vinylphenol, 4-vinylcatechol, and 4-vinylguaiacol, respectively. Among various ferulic acid decarboxylase enzymes, we chose the FADase from Enterobacter sp. Px6-4, whose crystal structure is known, and produced mutants to enhance its catalytic activity by random and site-directed mutagenesis. After three rounds of sequential mutations, FADase(F95L/D112N/V151I) showed approximately 34-fold higher catalytic activity than wild-type for the production of 4-vinylguaiacol from ferulic acid. Docking analyses suggested that the increased activity of FADase(F95L/D112N/V151I) could be due to formation of compact active site compared with that of the wild-type FADase. Considering the amount of phenolic compounds such as lignin monomers in the biomass components, successfully bioengineered FADase(F95L/D112N/V151I) from Enterobacter sp. Px6-4 could provide an ecofriendly biocatalytic tool for producing diverse styrene derivatives from biomass.

National pertussis surveillance in South Korea 1955-2011: epidemiological and clinical trends.

BACKGROUND: Although there has been substantial progress in controlling pertussis in South Korea, the reported number of pertussis case-patients has gradually been increasing during the last decade. To address this, we summarized the surveillance data on pertussis collected during the period 1955-2011. Detailed epidemiologic and clinical data were determined, primarily using data from recent years. METHODS: We analyzed data from the national surveillance system to describe the occurrence of pertussis. The annual numbers of reported pertussis case-patients were identified for the period 1955-2000. For 2001-2009, information including limited demographic characteristics and the date of onset of symptoms were identified. For 2010-2011, detailed epidemiologic and clinical information of reported pertussis case-patients were collected. RESULTS: During 1955-2011, the secular trend was characterized by a gradual decrease in the reported number of cases from 1955 to the late 1990s, then a recent increase starting in the early 2000s. In 2009, a large number of reported cases occurred in infants <1 year of age. In 2011, an increase in reported cases among adolescents and adults aged ≥15 years was observed. During 2010-2011, 29.8% of reported cases were not immunized and 11.3% had not been immunized in a timely manner. Of adolescents and adults aged ≥15 years, 91.7% did not have a record of immunization. CONCLUSIONS: During 2010-2011, a shift in age group was observed in pertussis case-patients: 33.8% were young infants <3 months of age and 29.0% were adolescents and adults ≥15 years of age. Considering that infants without timely vaccination may be vulnerable to an increased risk of pertussis infection, steps to provide timely vaccination to infants, to provide Tdap vaccination to adolescents and adults, and to enhance surveillance to capture adult pertussis cases should be taken in South Korea.