RESUMO
A sandwich electrochemiluminescence (ECL) immunosensor based on an N-(4-aminobutyl)-N-ethylisoluminol-graphene oxide-Ag nanoparticle (ABEI-GO-AgNPs) complex and cysteine silver nanowires (AgCysNWs) was prepared to detect prostate-specific antigen (PSA). Our results showed that an ECL signal probe, ABEI-GO-AgNPs, with an ultrahigh specific surface area, favorable catalytic properties, and electrical conductivity was prepared by a one-step synthesis method. ABEI-GO-AgNPs with good biocompatibility immobilized secondary antibody (Ab2) via AgN bonds. Furthermore, AgCysNWs containing many -COOH groups were prepared and used to enrich primary antibody (Ab1), which could be used as an affinity probe for the selective capture of PSA. Lastly, through layer-by-layer assembly, we established an ECL immunosensing platform for the sensitive detection of PSA. Under the optimized conditions, the designed ECL immunosensor showed promising sensitivity and selectivity for the detection of PSA in the linear range of 5.5 × 10-7-5.5 ng/mL, with a detection limit of 1.2 × 10-7 ng/mL. The constructed ECL sensing platform possessed good specificity, reproducibility, and stability and could detect PSA in actual human serum samples.
Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Nanofios , Técnicas Biossensoriais/métodos , Cisteína , Técnicas Eletroquímicas/métodos , Ouro/química , Humanos , Imunoensaio/métodos , Limite de Detecção , Medições Luminescentes/métodos , Luminol/análogos & derivados , Masculino , Nanopartículas Metálicas/química , Antígeno Prostático Específico , Reprodutibilidade dos Testes , Prata/químicaRESUMO
An ultrasensitive electrochemiluminescence based sandwich immunoassay is presented for determination of neuron specific enolase. The method uses silver-cysteine nanowires as the capture probe and a composite made of amino-modified reduced graphene oxide and nitrogen-doped carbon quantum dots as the signal probe. It was synthesized by covalent coupling of amino-modified reduced graphene oxide to the carboxy groups of nitrogen-doped carbon quantum dots. The nanowires possess a large specific surface and abundant functional groups which facilitate immobilizing the primary antibody (Ab1). The amino-modified reduced graphene oxide is employed as a carrier for loading a large number of the quantum dots and secondary antibody (Ab2). This increases the electrochemiluminescence intensity of quantum dots. Response to neuron specific enolase is linear in the 0.55 fg·mL-1 to 5.5 ng·mL-1 concentration range. It has a detection limit of 0.18 fg·mL-1 (at S/N = 3). The relative standard deviation (for n = 6) is less than 2.9%. The assay is highly sensitive, reproducible, selective and stable. Graphical abstractA novel electrochemiluminescence immunosensor is described that uses amino-modified reduced graphene oxide (amino-rGO), nitrogen-doped carbon quantum dots (N-CQDs) and silver-cysteine nanowires (SCNWs). It was applied to the determination of neuron specific enolase (NSE). Bovine serum albumin: BSA;1-ethyl-3-(3-dimethylaminopropyl)carbodiimide: (EDC;, N-hydroxysuccinimide: NHS.
Assuntos
Técnicas Eletroquímicas , Grafite/química , Imunoensaio , Medições Luminescentes , Fosfopiruvato Hidratase/sangue , Pontos Quânticos/química , Aminas/química , Técnicas Biossensoriais , Carbono/química , Humanos , Estrutura Molecular , Oxirredução , Tamanho da Partícula , Fosfopiruvato Hidratase/metabolismo , Propriedades de SuperfícieRESUMO
PURPOSE: Autism spectrum disorder (ASD) is a group of developmental brain disorders caused by genetic and environmental factors. The objective of this study was to investigate whether single nucleotide polymorphisms (SNPs) in genes related to immune function were associated with ASD in Chinese Han children. MATERIALS AND METHODS: A total of 201 children with ASD and 200 age- and gender-matched healthy controls were recruited from September 2012 to June 2106. A TaqMan probe-based approach was used to genotype SNPs corresponding to rs28532698 and rs4301112 in CD157, rs855867 in AIM2, and rs2237126 in JARID2. Case-control and case-only studies were performed to determine the contribution of SNPs to the predisposition of disease and its severity, respectively. RESULTS: Our results revealed that the genotypes and allele frequencies of these SNPs were not significantly associated with childhood ASD and its severity in this population. CONCLUSIONS: Results of our study suggest that these SNPs are not predictors of childhood ASD in the Chinese Han population. The discrepant results suggest the predictor roles of SNPs have to be determined in different ethnic populations due to genetic heterogeneity of ASD.
Assuntos
ADP-Ribosil Ciclase/genética , Antígenos CD/genética , Povo Asiático/genética , Transtorno do Espectro Autista/genética , Proteínas de Ligação a DNA/genética , Complexo Repressor Polycomb 2/genética , Polimorfismo de Nucleotídeo Único/genética , Adolescente , Povo Asiático/etnologia , Transtorno do Espectro Autista/diagnóstico , Transtorno do Espectro Autista/etnologia , Estudos de Casos e Controles , Criança , Pré-Escolar , Etnicidade/genética , Feminino , Proteínas Ligadas por GPI/genética , Frequência do Gene , Predisposição Genética para Doença/etnologia , Predisposição Genética para Doença/genética , Genótipo , Humanos , MasculinoRESUMO
Objective: To establish a fetal biparietal diameter (BPD)-gestational age formula based on the data of pregnant women from Xiaoshan District of Hangzhou, and to evaluate its application in prenatal screening. Methods: Data of 3500 pregnant women with gestational age between 15 weeks and 19 weeks+6 receiving prenatal screening in Xiaoshan Hospital during May 2014 and May 2015 were collected. BPDs were used to establish a localized BPD-gestational age formula. The localized formula was used to evaluate the prenatal screening risks in 1759 pregnant women with irregular menstrual cycles or uncertain last menstrual period (LMP) in Xiaoshan District, and the results were compared with those calculated using formula in LifeCycle 4.0. Results: With localized formula, the total positive rate of Down syndrome, trisomy 18 syndrome and deformity of neural tube was decreased from 6.96% to 5.85% ( P<0.05), in which the positive rate of Down syndrome decreased ( P<0.05), that of deformity of neural tube increased ( P<0.05), and that of trisomy 18 syndrome remained the same ( P>0.05). The median MoMs of free-hCG ß and α-fetoprotein calculated using localized formula were significantly different from those calculated using the formula in LifeCycle 4.0 (all P<0.05), and the former ones were more closer to 1. For women of fetus diagnosed with the above diseases, the positive rate calculated using localized formula was almost the same as that calculated using the formula in LifeCycle 4.0. Conclusion: BPD-gestational age formula should be localized based on the statistical analysis of the local population, which will help to reduce the false positive rate, and make the results more accurate and reliable in prenatal screening.
Assuntos
Pesos e Medidas Corporais/normas , Cefalometria/estatística & dados numéricos , Cefalometria/normas , Idade Gestacional , Cabeça/embriologia , Programas de Rastreamento/métodos , Programas de Rastreamento/normas , Diagnóstico Pré-Natal/estatística & dados numéricos , Diagnóstico Pré-Natal/normas , Adulto , Gonadotropina Coriônica Humana Subunidade beta/sangue , Gonadotropina Coriônica Humana Subunidade beta/normas , Cromossomos Humanos Par 18 , Síndrome de Down/diagnóstico , Síndrome de Down/embriologia , Medidas em Epidemiologia , Feminino , Desenvolvimento Fetal , Humanos , Programas de Rastreamento/estatística & dados numéricos , Ciclo Menstrual , Defeitos do Tubo Neural/diagnóstico , Defeitos do Tubo Neural/embriologia , Gravidez , Diagnóstico Pré-Natal/métodos , Valores de Referência , Trissomia/diagnóstico , Síndrome da Trissomía do Cromossomo 18 , alfa-Fetoproteínas/análise , alfa-Fetoproteínas/normasRESUMO
In electrospray ionization (ESI) quadrupole ion trap mass spectrometry, certain fragment ions generated in the ion trap from a series of sulfonic group containing ionic liquids (ILs) are found to undergo ion-molecule reactions with ESI solvent molecules (water, acetonitrile and methanol) to form adduct species. These unexpected solvated fragment ions severely complicate the interpretation of mass spectrometric data. Multi-stage fragmentation mass spectra were used to ascertain the chemical structures of these adduct species. It is important that solvation must be taken into account in order to prevent erroneous data interpretation for sulfonic group containing ILs.