Abstract
Diagnosis of seizure disorders such as epilepsy currently relies on clinical examination and electroencephalogram recordings and is associated with substantial mis-diagnosis. The miRNA, miR-134 (MIR134 in humans), has been found to be elevated in brain tissue after experimental status epilepticus and in human epilepsy cells and their detection in biofluids may serve as unique biomarkers. miRNAs from unprocessed human plasma and human cerebrospinal fluid samples were used in a novel electrochemical detection based on electrocatalytic platinum nanoparticles inside a centrifugal microfluidic device where the sandwich assay is formed using an event triggered release system, suitable for the rapid point-of-care detection of low abundance biomarkers of disease. The device has the advantage of controlling the rotation speed of the centrifugal device to pump nanoliter volumes of fluid at a set time and manipulate the transfer of liquids within the device. The centrifugal platform improves reaction rates and yields by proposing efficient mixing strategies to overcome diffusion-limited processes and improve mass transport rates, resulting in reduced hybridization times with a limit of detection of 1 pM target concentration. Plasma and cerebrospinal fluid samples (unprocessed) from patients with epilepsy or who experienced status epilepticus were tested and the catalytic response obtained was in range of the calibration plot. This study demonstrates a rapid and simple detection for epilepsy biomarkers in biofluid.
Highlights
Diagnosis of seizure disorders such as epilepsy currently relies on clinical examination and electroencephalogram recordings and is associated with substantial mis-diagnosis
An overflow system was implemented in the waste chambers ; this means that only when the second sample chamber (labelled (ii) in Fig. 1) has emptied into the waste chamber, it flows into the overflow part of the waste chamber (vii), which contains a control film (CF)
When this CF gets wet, it vents the pneumatic -channel (Lower channels, Layer 7) permitting the sample chamber (iii) to advance, and wet and dissolve the load film (LF). This liquid could flow into the electrode chamber. When this pneumatic chamber (iii) was vented, the liquid can only flow into the electrode chamber and not back through the venting channel into the waste; this was achieved through a physical barrier by extending the microchannels linking the CF and LF radially inward of the fluid in the sample chamber; this forces the fluid into the electrode chamber
Summary
Diagnosis of seizure disorders such as epilepsy currently relies on clinical examination and electroencephalogram recordings and is associated with substantial mis-diagnosis. The miRNA, miR-134 (MIR134 in humans), has been found to be elevated in brain tissue after experimental status epilepticus and in human epilepsy cells and their detection in biofluids may serve as unique biomarkers. The detection of one or more brain-specific miRNA in biofluids such as plasma or cerebrospinal fluid (CSF) may support diagnosis, predict seizures or guide treatment decisions for patients with epilepsy or status epilepticus[14]. Previous work[25] has shown that miR-134 can be detected at low levels using platinum nanoparticles (PtNPs) that are region-selectively decorated with probe strand nucleic acids complementary to miR-134 target in serum samples from epilepsy patients. The reported system required extraction of the miRNA from the biofluid before electrochemical detection was carried out These samples showed highly linear correlation in miR-134 measurement when compared to results obtained from Taqman-based PCR25. We demonstrate the detection of miRNA in an unprocessed biofluid samples as a routine detection strategy in a clinical setting, in a microfluidic device
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