Rapid Analysis for Staphylococcus aureus via Microchip Capillary Electrophoresis

Abstract

Staphylococcus aureus (S. aureus) is one of the most common pathogens for nosocomial and community infections, which is closely related to the occurrence of pyogenic and toxic diseases in human beings. In the current study, a lab-built microchip capillary electrophoresis (microchip CE) system was employed for the rapid determination of S. aureus, while a simple-to-use space domain internal standard (SDIS) method was carried out for the reliable quantitative analysis. The precision, accuracy, and reliability of SDIS were investigated in detail. Noted that these properties could be elevated in SDIS compared with traditional IS method. Remarkably, the PCR products of S. aureusnuc gene could be identified and quantitated within 80 s. The theoretical detection limit could achieve a value of 0.066 ng/μL, determined by the using SDIS method. The current work may provide a promising detection strategy for the high-speed and highly efficient analysis of pathogens in the fields of food safety and clinical diagnosis.

Rapid Detection of Staphylococcus aureus Using Paper-Derived Electrochemical Biosensors

“…While a number of technologies have already been presented for the detection of S. aureus, − including sensors based on aptamers, − immunoglobulins, , proteases, or PCR amplification, the proposed sensor could not only avoid the use of biorecognition elements but also sample pretreatments, as it could be directly applied to the skin.…”

“…Specifically, although the proposed approach would enable identifying an infection (yes/no answer) within 20 min, the assay can be followed for longer periods of time (additional 40 min) to render a signal that is proportional to the initial density of bacteria (Supporting Information Figure S11) and that can detect down to 0.1 CFU/mL. While a number of technologies have already been presented for the detection of S. aureus, 86−89 including sensors based on aptamers, 90−92 immunoglobulins, 93,94 proteases, 49 or PCR amplification, 95 the proposed sensor could not only avoid the use of biorecognition elements but also sample pretreatments, as it could be directly applied to the skin.…”

Ultradense Electrochemical Chips with Arrays of Nanostructured Microelectrodes to Enable Sensitive Diffusion-Limited Bioassays

“…Briefly, the respiratory activity of the bacteria involves the generation of electron-donor enzymes that promote the reduction of hydrophilic electron-acceptor species such as [Fe(CN) 6 ] 3– , thus forming [Fe(CN) 6 ] 4– that can serve as a marker of the concentration of bacterium . Based on this process, amperometric sensors have been addressed for the fast and simple quantification of bacteria by collecting the anodic currents derived from [Fe(CN) 6 ] 4– oxidation, which can be related to the number of respiring cells. , Such CR 4 -sensing methods entail advantages over labor-intensive and time-consuming traditional techniques (e.g., broth dilution, agar disk diffusion, and antimicrobial gradient assay) and biosensors − by combining the merits of electrochemical platforms (speed, simplicity, low cost, and portability) with the absence of recognition elements. Overall, these compounds demand complex, expensive, and low-yield in vitro synthesis, and they are prone to denaturation, thus reducing the shelf life of the biosensing device (e.g., a few days when stored at 4.0 °C in buffer). , …”

Recent advances in microchip electrophoresis for analysis of pathogenic bacteria and viruses

“…The method involved PCR amplification of the nuc gene of S. aureus , and the PCR products were labelled with SYBR Green I. Then, the fluorescently labelled PCR products of the nuc gene of S. aureus bacteria were separated and detected by ME within only 80 s [109]. The drawback of using SYBR Green I dye is its tendency to bind to any dsDNA sequences, generating false‐positive results.…”