RESUMO
A very sensitive and selective colorimetric biosensor for the measurement of mercury ion (Hg2+) in environmental samples has been developed using functionalized gold nanoparticles with bromelain enzyme (brn-AuNPs). This work has shown that Hg2+ measurement based on spectrophotometer and digital image analysis is a very innovative and successful method for providing an effective preliminary system and has promise for the future of water quality biomonitoring. Response Surface Methodology (RSM), a Box-Behnken design-based technique, was used to identify the optimum levels of functionalization of bromelain to AuNPs. The created model's validity was confirmed, and statistical analysis revealed that the ideal functionalize conditions were 1 mM of AuNPs, functionalize with 0.59 mM bromelain concentration on 14 â temperature and 72 h incubation time. The lowest colorimetric detection concentration (LOD) of brn-AuNPs of Hg2+ was 0.0092 ppm and 0.011 ppm for spectrophotometer and digital image analysis. As shown, digital image analysis had advantages based on the LOD result comparable to UV-VIS spectrophotometer. The practical application of the brn-AuNPs sensing was proven with mercury determination in water samples. The present study developed a robust sensor, which successfully implemented in a compact portable sensor kit, turning this sensor into a very potent tool for the development water quality biomonitoring system of Hg2+ application.
RESUMO
In this study, an electrochemical DNA biosensor was developed based on the fabrication of silicon nanowires/platinum nanoparticles (SiNWs/PtNPs) on a screen-printed carbon electrode (SPCE) for the detection of Sus scrofa mitochondrial DNA (mtDNA) in food utilizing a new hybrid indicator, ferrocenylnaphthalene diimide (FND). The morphology and elemental composition of the SiNWs/PtNPs-modified SPCE was analyzed by field emission scanning electron microscopy (FESEM) combined with energy dispersive X-ray spectroscopy (EDX). Cyclic voltammetry (CV) was used to study the electrical contact between the PtNPs and the screen-printed working electrode through SiNWs, while electrochemical impedance spectroscopy (EIS) was used to measure the charge transfer resistance of the modified electrode. The results clearly showed that the SiNWs/PtNPs were successfully coated onto the electrode and the effective surface area for the SiNWs/PtNPs-modified SPCE was increased 16.8 times as compared with that of the bare SPCE. Differential pulse voltammetry used for the detection of porcine DNA with FND as an intercalator confirmed its specific binding to the double-stranded DNA (dsDNA) sequences. The developed biosensor showed a selective response towards complementary target DNA and was able to distinguish non-complementary and mismatched DNA oligonucleotides. The SiNWs/PtNPs-modified SPCE that was fortified with DNA hybridization demonstrated good linearity in the range of 3 × 10-9 M to 3 × 10-5 M (R 2 = 0.96) with a detection limit of 2.4 × 10-9 M. A cross-reactivity study against various types of meat and processed food showed good reliability for porcine samples.
RESUMO
A rapid and sensitive sandwich electrochemical immunosensor was developed based on the fabrication of the graphene/polyaniline (GP/PANI) nanocomposite onto screen-printed gold electrode (SPGE) for detection of tuberculosis biomarker 10-kDa culture filtrate protein (CFP10). The prepared GP/PANI nanocomposite was characterized using Fourier transform infrared spectroscopy (FTIR) and field emission scanning electron microscopy (FESEM). The chemical bonding and morphology of GP/PANI-modified SPGE were studied by Raman spectroscopy and FESEM coupled with energy dispersive X-ray spectroscopy, respectively. From both studies, it clearly showed that GP/PANI was successfully coated onto SPGE through drop cast technique. Cyclic voltammetry was used to study the electrochemical properties of the modified electrode. The effective surface area for GP/PANI-modified SPGE was enhanced about five times compared with bare SPGE. Differential pulse voltammetry was used to detect the CFP10 antigen. The GP/PANI-modified SPGE that was fortified with sandwich type immunosensor exhibited a wide linear range (20â»100 ng/mL) with a low detection limit of 15 ng/mL. This proposed electrochemical immunosensor is sensitive, low sample volume, rapid and disposable, which is suitable for tuberculosis detection in real samples.
Assuntos
Técnicas Biossensoriais , Fragmentos de Peptídeos/isolamento & purificação , Tuberculose/diagnóstico , Compostos de Anilina/química , Diagnóstico Precoce , Técnicas Eletroquímicas/métodos , Grafite/química , Humanos , Limite de Detecção , Nanopartículas Metálicas/química , Microscopia Eletrônica de Varredura/métodos , Mycobacterium tuberculosis/isolamento & purificação , Mycobacterium tuberculosis/patogenicidade , Fragmentos de Peptídeos/imunologia , Espectroscopia de Infravermelho com Transformada de Fourier , Análise Espectral Raman , Tuberculose/imunologiaRESUMO
Tuberculosis (TB) remains one of the most devastating infectious diseases and its treatment efficiency is majorly influenced by the stage at which infection with the TB bacterium is diagnosed. The available methods for TB diagnosis are either time consuming, costly or not efficient. This study employs a signal generation mechanism for biosensing, known as Plasmonic ELISA, and computational intelligence to facilitate automatic diagnosis of TB. Plasmonic ELISA enables the detection of a few molecules of analyte by the incorporation of smart nanomaterials for better sensitivity of the developed detection system. The computational system uses k-means clustering and thresholding for image segmentation. This paper presents the results of the classification performance of the Plasmonic ELISA imaging data by using various types of classifiers. The five-fold cross-validation results show high accuracy rate (>97%) in classifying TB images using the entire data set. Future work will focus on developing an intelligent mobile-enabled expert system to diagnose TB in real-time. The intelligent system will be clinically validated and tested in collaboration with healthcare providers in Malaysia.