查看更多>>摘要：Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most devastating viruses in the swine industry and causes major economic losses. To date, there has not been an effective antiviral treatment for the disease. We have shown in previous studies that culture supernatant of Actinobacillus pleuropneumoniae (App), the causative agent of porcine pleuropneumonia, possesses antiviral activity in vitro against PRRSV, and we have clearly established that the antiviral activity was mediated by small molecular weight (i.e., <1 kDa), heat resistant metabolites present in the App supernatant ultrafiltrates. However, the identity of those metabolites remains unknown. The objective of the current study was to identify the active metabolites using untargeted and targeted mass spectrometry-based metabolomics and test their respective antiviral activity against PRRSV in the Jude Porcine Lung Epithelial Cell Line (SJPL). The results presented reveal very significant antiviral activity of App supernatant ultrafiltrates against PRRSV in SJPL cells. Consequently, we identified and quantified several adenosine nucleotide metabolites present in App supernatant ultrafiltrates using mass spectrometry-based metabolomics, and the concentrations detected were very high. SJPL cells infected with PRRSV and treated with 2 '-adenosine monophosphate (2-AMP), 3 '-adenosine monophosphate (3-AMP) or 5 '-adenosine monophosphate (5-AMP) significantly reduced PRRSV infection. Interestingly, many antiviral drugs or prodrugs are adenosine analogs, and the mechanism of action was previously elucidated. Currently marketed nucleoside analog drugs could potentially be used to treat PRRSV infection.
查看更多>>摘要：Ratiometric electrochemical aptasensors based on the two different redox reporters can hardly achieve the efficient self-calibration, as a result of variations between reporters. In this work, we reported a ratiometric electrochemical aptasensor using ferrocene as a single redox reporter (Fc) to achieve self-calibrating electro-chemical detection of aflatoxin B1 (AFB1). The hybrid duplex of Fc-labeled AFB1 aptamer (Fc-Apt) and Fc-labeled assistant DNA (Fc-aDNA) was designed to construct the sensing interface. Such DNA structure adopted the fixed molar ratio (1:1) and locations (the distances to electrode) of these two types of Fc (i.e., Fc-Apt and Fc-aDNA). In this way, the ratio of redox currents from Fc-Apt (IFc-Apt) and Fc-aDNA (IFc-aDNA) kept stable under the varied temperatures (15-40 degrees C) and pH values (4-10). Moreover, values of IFc-Apt and IFc-aDNA under the different conditions can be calculated simply by recording the total current of Fc (IFc-total). For analysis, the recognition of AFB1 by Fc-Apt led to the striping of their complex from the aptasensor, resulting in a decrease in IFc-Apt while IFc-aDNA remained stable. Consequently, the developed aptasensor using the ratio of IFc-Apt/IFc-aDNA as a yardstick offered a linear range of 0.1-10000 pg mL(-1) with a detection limit of 0.012 pg mL(-1) for the detection of AFB1. The applicability of aptasensor was validated by corn sample analysis, which exhibited comparable reliability and accuracy of gold standard method, i.e., HPLC-MS/MS. Our work provided an efficient strategy to fabricate high-performance ratiometric electrochemical aptasensors.
查看更多>>摘要：The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been spreading worldwide for more than a year and has undergone several mutations and evolutions. Due to the lack of effective therapeutics and long active vaccines, accurate and large-scale screening and early diagnosis of infected individuals are crucial to control the pandemic. Nevertheless, the current widely used RT-qPCR-based methods suffer from complicated temperature control, long processing time and the risk of false-negative results. Herein, we present a three-way junction induced exponential rolling circle amplification (3WJ-eRCA) combined MALDI-TOF MS assay for SARSCoV-2 detection. The assay can detect simultaneously the target nucleocapsid (N) and open reading frame 1 ab (orf1ab) genes of SARS-CoV-2 in a single test within 30 min, with an isothermal process (55 degrees C). High specificity to discriminate SARS-CoV-2 from other coronaviruses, like SARS-CoV, MERS-CoV and bat SARS-like coronavirus (bat-SL-CoVZC45), was observed. We have further used the method to detect pseudovirus of SARS-CoV-2 in various matrices, e.g. water, saliva and urine. The results demonstrated a great potential of the method for large scale screening of COVID-19, which is an important part of the pandemic control.
查看更多>>摘要：The rapid and simple synthesis of highly efficient multi-emission luminescent metal-organic frameworks (LMOFs) with down/up-conversion fluorescence properties becomes critical in potential optical sensing. Here, we present a luminescent dual-ligand co-assisted strategy to one-pot prepare LMOFs for multiplex fluorescence sensing using carbon dots (CDs) and 2-methylimidazole (2-MIM) as dual-ligand to induce Zn2+ on zeolitic imidazolate framework-8 surface (CDs/MIM-Zn). The CDs/MIM-Zn exhibited efficient fluorescence-enhanced dual channels triple-emission with Stokes and anti-Stokes type excitation profiles. With the assistance of Stokes and anti-Stokes type optical property of CDs and the intrinsic porosity of MOFs structure, a rapid and sensitive dual-channel detection excited at 370 nm and 790 nm was performed to fluorescence turn off-on detect metal ions and reduced glutathione in real blood samples with the satisfactory recoveries of 98.3-106.4%. The dual channels triple-emission strategy ensured multiplex detection of more analytes in parallel.
NSTL
Elsevier