ABSTRACT
Sepsis is a life-threatening condition, which is irreversible if diagnosis and intervention are delayed. The response of the immune cells towards an infection triggers widespread inflammation through the production of cytokines, which may result in multiple organ dysfunction and eventual death. Conventional detection techniques fail to provide a rapid diagnosis because of their limited sensitivity and tedious protocol. This study proposes a point-of-care (POC) electrochemical biosensor that overcomes the limitations of current biosensing technologies in the clinical setting by its integration with electrokinetics, enhancing the sensitivity to picogram level compared with the nanogram limit of current diagnostic technologies. This biosensor promotes the use of a microelectrode strip to address the limitations of conventional photolithographic fabrication methods. Tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and microRNA-155 (miR-155) were monitored in a lipopolysaccharide (LPS)-induced septic mouse model. The optimum target hybridization time in a high conductivity medium was observed to be 60 s leading to the completion of the whole operation within 5 min compared with the 4-h detection time of the traditional enzyme-linked immunosorbent assay (ELISA). The limit of detection (LOD) was calculated to be 0.84, 0.18, and 0.0014 pg mL-1, respectively. This novel sensor may have potential for the early diagnosis of sepsis in the clinical setting.
Subject(s)
Biosensing Techniques , MicroRNAs , Sepsis , Mice , Animals , Lipopolysaccharides/toxicity , Point-of-Care Systems , Disease Models, Animal , Biosensing Techniques/methods , Sepsis/chemically induced , Sepsis/diagnosis , Biomarkers/analysis , Tumor Necrosis Factor-alpha , MicroRNAs/analysisABSTRACT
Surface plasmon resonance (SPR) has emerged as one of the most efficient and attractive techniques for optical sensors in biological applications. The traditional approach of an EC (electrochemical)-SPR biosensor to generate SPR is by adopting a prism underneath the sensing substrate, and an angular scan is performed to characterize the reflectivity of target analytes. In this paper, we designed and investigated a novel optical biosensor based on a hybrid plasmonic and electrochemical phenomenon. The SPR was generated from a thin layer of gold nanohole array on a glass substrate. Using C-Reactive Protein (CRP) as the target analyte, we tested our device for different concentrations and observed the optical response under various voltage bias conditions. We observed that SPR response is concentration-dependent and can be modulated by varying DC voltages or AC bias frequencies. For CRP concentrations ranging from 1 to 1000 µg/mL, at the applied voltage of -600â mV, we obtained a limit of detection for this device of 16.5â ng/mL at the resonance peak wavelength of 690â nm. The phenomenon is due to spatial re-distribution of electron concentration at the metal-solution interface. The results suggest that CRP concentration can be determined from the SPR peak wavelength shift by scanning the voltages. The proposed new sensor structure is permissible for various future optoelectronic integration for plasmonic and electrochemical sensing.
ABSTRACT
Traditional immunosensors are often limited by low sensitivity and long detection times, for they usually depend on passive diffusion-dominated transport of target analytes for the binding reaction with a bio-recognition element such as enzymes, antibodies, and aptamers. Numerous studies rely on electric field manipulation by using alternating current (AC) electrokinetics to enhance the hybridization rate and reduce the hybridization time for faster and more efficient detection. This study demonstrated a rapid electrochemical aptasensor integrated with an AC electroosmotic (ACEO) flow phenomenon for the enhanced target hybridization of microRNA-155 (miR-155). Optimization of the electrokinetic conditions for target collection resulted in a saturation point after 75 s miR-155 was detected within the range of 1 aM-10 pM with a detection limit of 1 aM, which is 100 times lower and about 50 times faster compared with the conventional diffusion-dependent detection done for 1 h. The detection was also done in spiked serum samples, and a concentration range within the required detection range was obtained. The highly sensitive and specific results allow for the rapid and real-time sensing of target biomarkers, which can be used for the early detection of infection.
Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , MicroRNAs , Electrochemical Techniques , Electroosmosis , Immunoassay , Limit of Detection , Nucleic Acid HybridizationABSTRACT
In this study, the time-dependent reaction between 11-mercaptoundecanoic acid (11-MUA) and 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide/N-hydroxysuccinimide (EDC/NHS) is precisely characterized using surface enhanced infrared absorption spectroscopy (SEIRAS). According to the high correlation between the spectral results of SEIRAS and the electrochemical behavior, it strongly demonstrates that the EDC/NHS reaction would be obviously interfered by phosphate ions in the neutral pH condition (pH = 7.0).