RESUMO
A signal-amplified platform was designed to construct a label-free electrochemical aptasensor for lead ions (Pb2+) assay. First, flower-like molybdenum disulfide-supported AuNPs (AuNPs@MoS2) nanocomposites were synthesized and used as substrates for modifying the electrode. The AuNPs@MoS2 material possessed large surface area and superior biocompatibility, which was beneficial to improve the loading amount of the complementary DNA (cDNA) and amplified the response signal. Importantly, the prepared core-shell Pt@Pd bimetallic nanoparticles (Pt@PdNPs) were used to conjugate with redox marker thionine (Thi) and aptamer (Apt) for further signal amplification; the obtained signal probes (Thi-Pt@PdNPs-Apt) were connected by the cDNA assembled on the electrode through DNA hybridization. Differential pulse voltammetry was performed to monitor the signal of Thi. After incubating of aptasensor with Pb2+, the specific recognition of Pb2+ and Apt resulted in the dissociation of aptamer-cDNA complex, thereby the Thi-Pt@PdNPs-Apt separated from the electrode surface and decreased current response was obtained. The prepared electrochemical sensor exhibited linear response to Pb2+ in the range 5.0 × 10-4-100 nM and a detection limit of 1.0 × 10-4 nM was achieved. The sensor was applied to the determination of Pb2+ in actual sample with high sensitivity and accuracy, demonstrating potential applications in heavy metal monitoring.
RESUMO
A novel electrochemiluminescence (ECL) aptasensor, using graphite carbonitride (g-C3N4) capped CdS quantum dots (CdS QDs@g-C3N4) and Au nanoparticles decorated triethanolamine (AuNPs@TEOA) as dual coreactants, was proposed for the determination of mucin 1 (MUC1). Higher ECL efficiency was acquired due to the double enhancement contribution of CdS QDs and TEOA to Ru (bpy)32+ ECL. Additionally, AuNPs@TEOA also acted as nanocarrier for MUC1 aptamer immobilization. After the aptasensor was incubated in target MUC1, the decreased ECL emission was obtained because of the poor conductivity of MUC1. The ECL aptasensor displayed a good linear correlation for MUC1 in the range 0.1 pg mL-1 -1000 ng mL-1, and the detection limit was 33 fg mL-1. MUC1 spiked into human serum samples was quantified to assess the practicability of the ECL aptasensor.
Assuntos
Ouro , Nanopartículas Metálicas , Humanos , Mucina-1 , Técnicas Eletroquímicas , Medições LuminescentesRESUMO
A novel electrochemiluminescence (ECL) aptasensor for the determination of lincomycin (LIN) was developed based on CdS QDs/carboxylated g-C3N4 (CdS QDs/C-g-C3N4). CdS QDs/C-g-C3N4 served as the substrate of the aptasensor, and then CdS QDs/C-g-C3N4-modified electrode was incubated with aptamer DNA (Apt-DNA). When the non-specific sites of the electrode surface was blocked by 6-mercaptohexanol, the ferrocene-labeled probe (Fer-DNA) was assembled onto the electrode surface through base complementation with Apt-DNA. In the absence of LIN, the ECL signal was quenched effectively by Fer-DNA and a decreased ECL emission (off state) was acquired. On the contrary, LIN was specifically bond with Apt-DNA, and Fer-DNA was detached from the aptasensor surface because of the deformation of Apt-DNA, resulting in an effectively enhanced ECL signal (on state). The constructed ECL aptasensor exhibited a wide detection range for LIN determination (0.05 ng mL-1-100 µg mL-1) with a low detection limit (0.02 ng mL-1). Importantly, the proposed ECL aptasensor showed outstanding accuracy and specificity for LIN determination, and also provided a potential strategy for other antibiotic determinations.
Assuntos
DNA , LincomicinaRESUMO
Cyanidin-3-O-glucoside (C3G), a natural antioxidant, plays multiple physiological or pathological roles in maintaining human health; thereby, designing advanced sensors to achieve specific recognition and high-sensitivity detection of C3G is significant. Herein, an imprinted-type electrochemiluminescence (ECL) sensing platform was developed using core-shell Ru@SiO2-CMIPs, which were prepared by covalent organic framework (COF)-based molecularly imprinted polymers (CMIPs) embedded in luminescent Ru@SiO2 cores. The C3G-imprinted COF shell not only helps generate a steady-enhanced ECL signal, but also enables specific recognition of C3G. When C3G is bound to Ru@SiO2-CMIPs with abundant imprinted cavities, resonance energy transfer (RET) behavior is triggered, resulting in a quenched ECL response. The constructed Ru@SiO2-CMIPs nanoprobes exhibit ultra-high sensitivity, absolute specificity, and an ultra-low detection limit (0.15 pg mL-1) for analyzing C3G in food matrices. This study provides a means to construct an efficient and reliable molecular imprinting-based ECL sensor for food analysis.
Assuntos
Antocianinas , Técnicas Eletroquímicas , Glucosídeos , Medições Luminescentes , Estruturas Metalorgânicas , Impressão Molecular , Rutênio , Dióxido de Silício , Antocianinas/química , Antocianinas/análise , Dióxido de Silício/química , Medições Luminescentes/métodos , Técnicas Eletroquímicas/métodos , Rutênio/química , Glucosídeos/química , Glucosídeos/análise , Estruturas Metalorgânicas/química , Limite de Detecção , Polímeros Molecularmente Impressos/químicaRESUMO
A novel global search algorithm based method is proposed to separate MR images blindly in this paper. The key point of the method is the formulation of the new matrix which forms a generalized permutation of the original mixing matrix. Since the lowest entropy is closely associated with the smooth degree of source images, blind image separation can be formulated to an entropy minimization problem by using the property that most of neighbor pixels are smooth. A new dataset can be obtained by multiplying the mixed matrix by the inverse of the new matrix. Thus, the search technique is used to searching for the lowest entropy values of the new data. Accordingly, the separation weight vector associated with the lowest entropy values can be obtained. Compared with the conventional independent component analysis (ICA), the original signals in the proposed algorithm are not required to be independent. Simulation results on MR images are employed to further show the advantages of the proposed method.
Assuntos
Processamento de Imagem Assistida por Computador/métodos , Imageamento por Ressonância Magnética/métodos , Algoritmos , Encéfalo/patologia , Mapeamento Encefálico/métodos , Simulação por Computador , Entropia , Humanos , Modelos Lineares , Análise de Componente Principal , Processamento de Sinais Assistido por ComputadorRESUMO
A high-mortality swine disease, the highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS), reappeared in some regions of China in 2009. To explore the possible mechanisms underlying the emergence of HP-PRRSV and more fully understand the extent of the genetic diversity of this virus in China, the complete genome of 14 isolates from 10 provinces in China from 2009 were analyzed. Full-length genome sequencing analysis showed that the 14 isolates were closely related to HP-PRRSV, with 98.0-98.9% nucleotide similarity, although 2 of the 14 strains exhibited a new, discontinuous 29-amino acid deletion in the Nsp2 gene. Furthermore, amino acid analysis of the GP5 protein indicated that the 14 isolates had a concurrent mutation in a decoy epitope and different mutations in glycosylation sites. Additionally, the antigenic drift in GP3 and a 1-nucleotide deletion in both the 5'-UTR and 3'-UTR, which are found in almost all highly pathogenic Chinese PRRSV isolates, were examined in all 14 isolates. The phylogenetic analysis showed that the 14 strains belonged to the North American genotype and were clustered in a subgroup with other HP-PRRSV isolates that have been found in China since 2006. However, compared with other Chinese HP-PRRSV isolates collected in 2006-2008, the phylogenetic tree showed that the 14 isolates had a closer relationship with each other. These results indicated that HP-PRRSV remained an extensive pandemic, affecting swine farms in China in 2009 and revealed new genetic diversity.
Assuntos
Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Sequência de Aminoácidos , Animais , China/epidemiologia , Variação Genética , Genoma Viral , Dados de Sequência Molecular , Filogenia , Síndrome Respiratória e Reprodutiva Suína/epidemiologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/classificação , Vírus da Síndrome Respiratória e Reprodutiva Suína/isolamento & purificação , Alinhamento de Sequência , Suínos , Regiões não Traduzidas , Proteínas do Envelope Viral/química , Proteínas do Envelope Viral/genética , Proteínas Estruturais Virais/química , Proteínas Estruturais Virais/genéticaRESUMO
The Thiverval vaccine strain of classical swine fever virus (CSFV) was derived from virulent Alfort strain through the serial passages in cells at 29-30 degrees C. In this study, we determined the complete genome sequence of this strain and found that its genome contains one open reading frame (ORF) that encodes a polyprotein with 3,898 amino acids. The 5'-UTR of Thiverval is 373 nt long with only one mutation at position 220. In contrast, the length of 3'-UTR is highly heterogeneous ranging from 233 to 259 bp. The heterogeneity of length of the 3'-UTR was due to an insertion of a variable length of T-rich sequence ranging from 6 to 32 nt. The insertion may change the structure and free energy of the 3'-UTR, resulting in a destabilization of the 3'-UTR. Sequence alignment of Thiverval and other CSFV strains showed 85.2-99.6% identities at the nucleotide level and 92.5-99.5% at the amino acid level. The phylogenetic tree analysis of the complete ORF, partial region of E2, and NS5B suggests that the CSFV Thiverval strain belongs to genetic group 1 and subgroup 1.1. The results from this study provide insight into the molecular mechanism of the attenuation of Thiverval vaccine strain.