scholarly journals Analytical Molecular Diagnosis of Cervical Cancer via Paper Microfluidic Chip

Proceedings ◽  
2018 ◽  
Vol 2 (25) ◽  
pp. 1556
Author(s):  
Melike Karakaya

In this work, we focused on the development of a novel point of care (POC) paper-based analytical microfluidic chip. The system has been developed for paper-based extraction, non-enzymatic amplification and electroanalytical detection of human papillomavirus (HPV). The device comprises paper extraction support material as well as paper-based detection tool. Herein, a special DNA modification method has been utilized to allow non-enzymatic amplification by using microbeads and silver nanoparticles labeled primers. The device is capable of extracting more than 10–100 copies per mL of DNA approximately in 15 min along with a single step extraction process from patient samples. In addition, it is able to detect low concentrations taking just less than 10 min with high selectivity to HPV kinds of 16&18. It only takes the low-cost point of care device less than 40 min with a low limit of detection (LOD) to complete the whole process.

Sensors ◽  
2021 ◽  
Vol 21 (12) ◽  
pp. 3985
Author(s):  
Nan Wan ◽  
Yu Jiang ◽  
Jiamei Huang ◽  
Rania Oueslati ◽  
Shigetoshi Eda ◽  
...  

A sensitive and efficient method for microRNAs (miRNAs) detection is strongly desired by clinicians and, in recent years, the search for such a method has drawn much attention. There has been significant interest in using miRNA as biomarkers for multiple diseases and conditions in clinical diagnostics. Presently, most miRNA detection methods suffer from drawbacks, e.g., low sensitivity, long assay time, expensive equipment, trained personnel, or unsuitability for point-of-care. New methodologies are needed to overcome these limitations to allow rapid, sensitive, low-cost, easy-to-use, and portable methods for miRNA detection at the point of care. In this work, to overcome these shortcomings, we integrated capacitive sensing and alternating current electrokinetic effects to detect specific miRNA-16b molecules, as a model, with the limit of detection reaching 1.0 femto molar (fM) levels. The specificity of the sensor was verified by testing miRNA-25, which has the same length as miRNA-16b. The sensor we developed demonstrated significant improvements in sensitivity, response time and cost over other miRNA detection methods, and has application potential at point-of-care.


Biosensors ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 4
Author(s):  
Donggee Rho ◽  
Seunghyun Kim

An optical cavity-based biosensor (OCB) has been developed for point-of-care (POC) applications. This label-free biosensor employs low-cost components and simple fabrication processes to lower the overall cost while achieving high sensitivity using a differential detection method. To experimentally demonstrate its limit of detection (LOD), we conducted biosensing experiments with streptavidin and C-reactive protein (CRP). The optical cavity structure was optimized further for better sensitivity and easier fluid control. We utilized the polymer swelling property to fine-tune the optical cavity width, which significantly improved the success rate to produce measurable samples. Four different concentrations of streptavidin were tested in triplicate, and the LOD of the OCB was determined to be 1.35 nM. The OCB also successfully detected three different concentrations of human CRP using biotinylated CRP antibody. The LOD for CRP detection was 377 pM. All measurements were done using a small sample volume of 15 µL within 30 min. By reducing the sensing area, improving the functionalization and passivation processes, and increasing the sample volume, the LOD of the OCB are estimated to be reduced further to the femto-molar range. Overall, the demonstrated capability of the OCB in the present work shows great potential to be used as a promising POC biosensor.


2021 ◽  
Author(s):  
Binfeng Yin ◽  
Xinhua Wan ◽  
Mingzhu Yang ◽  
Changcheng Qian ◽  
A S M Muhtasim Fuad Sohan

Abstract Background: Simultaneous and timely detection of C-reactive protein (CRP), procalcitonin (PCT), and interleukin-6 (IL-6) provides effective information for the accurate diagnosis of infections. Early diagnosis and classification of infections increase the cure rate while decreasing complications, which is significant for severe infections, especially for war surgery. However, traditional methods rely on laborious operations and bulky devices. On the other hand, point-of-care (POC) methods suffer from limited robustness and accuracy. Therefore, it is of urgent demand to develop POC devices for rapid and accurate diagnosis of infections to fulfill on-site militarized requirements.Methods: We developed a wave-shaped microfluidic chip (WMC) assisted multiplexed detection platform (WMC-MDP). WMC-MDP reduces detection time and improves repeatability through premixing of the samples and reaction of the reagents. We further combined the detection platform with the streptavidin-biotin (SA-B) amplified system to enhance the sensitivity while using chemiluminescence (CL) intensity as signal readout. We realized simultaneous detection of CRP, PCT, and IL-6 on the detection platform and evaluated the sensitivity, linear range, selectivity, and repeatability. Finally, we finished detecting 15 samples from volunteers and compared the results with commercial ELISA kits.Results: Detection of CRP, PCT, and IL-6 exhibited good linear relationships between CL intensities and concentrations in the range of 1.25-40 μg/mL, 0.4-12.8 ng/mL, and 50-1600 pg/mL. The limit of detection (LOD) of CRP, PCT, and IL-6 were 0.54 μg/mL, 0.11 ng/mL, and 16.25 pg/mL, respectively. WMC-MDP is capable of good adequate selectivity and repeatability. The whole detection procedure takes only 22 minutes that meets the requirements of a POC device. Results of 15 samples from volunteers were consistent with the results detected by commercial ELISA kits.Conclusion: WMC-MDP allows simultaneous, rapid, and sensitive detection of CRP, PCT, and IL-6 with satisfactory selectivity and repeatability, requiring minimal manipulation. However, WMC-MDP takes advantage of being a microfluidic device showing the coefficients of variation less than 10% enabling WMC-MDP to be a type of POCT. Therefore, WMC-MDP provides a promising alternative to point-of-care testing (POCT) of multiple biomarkers. We believe the practical application of WMC-MDP in militarized fields will revolutionize infection diagnosis for soldiers.


2011 ◽  
Vol 57 (5) ◽  
pp. 753-761 ◽  
Author(s):  
Ulrich Y Schaff ◽  
Greg J Sommer

BACKGROUND Centrifugal “lab on a disk” microfluidics is a promising avenue for developing portable, low-cost, automated immunoassays. However, the necessity of incorporating multiple wash steps results in complicated designs that increase the time and sample/reagent volumes needed to run assays and raises the probability of errors. We present proof of principle for a disk-based microfluidic immunoassay technique that processes blood samples without conventional wash steps. METHODS Microfluidic disks were fabricated from layers of patterned, double-sided tape and polymer sheets. Sample was mixed on-disk with assay capture beads and labeling antibodies. Following incubation, the assay beads were physically separated from the blood cells, plasma, and unbound label by centrifugation through a density medium. A signal-laden pellet formed at the periphery of the disk was analyzed to quantify concentration of the target analyte. RESULTS To demonstrate this technique, the inflammation biomarkers C-reactive protein and interleukin-6 were measured from spiked mouse plasma and human whole blood samples. On-disk processing (mixing, labeling, and separation) facilitated direct assays on 1-μL samples with a 15-min sample-to-answer time, <100 pmol/L limit of detection, and 10% CV. We also used a unique single-channel multiplexing technique based on the sedimentation rate of different size or density bead populations. CONCLUSIONS This portable microfluidic system is a promising method for rapid, inexpensive, and automated detection of multiple analytes directly from a drop of blood in a point-of-care setting.


2016 ◽  
Vol 2 (3_suppl) ◽  
pp. 14s-14s
Author(s):  
Benjamin A. Katchman ◽  
Joseph T. Smith ◽  
Jennifer Blain Christen ◽  
Karen S. Anderson

Abstract 62 One of the key roadblocks limiting the transition of high-sensitivity and high-specificity point-of-care technologies from the research laboratory to wide spread use is the availability of a low-cost-high-volume manufacturing technology. This work presents a new interdisciplinary approach combining low cost commercial display manufacturing technology with programmable high density protein microarray printing technology to fabricate disposable point-of-care immunosensors with clinical level sensitivity. Our approach is designed to leverage advances in commercial display technology to reduce pre-functionalized biosensor substrate costs to pennies per cm2, as well as to leverage the display industry’s ability to manufacture an immense number of low cost consumer electronic products annually. For this work, we demonstrate that our new approach can offer diagnostic sensitivity at or below 10 pg/mL, which approaches the lower limit of detection of typical clinical laboratory instrumentation. Our new approach is also designed to overcome the limited analytical sensitivity of existing POC devices (>100x improved sensitivity). It also contains new capability for multiplexed biomarker detection (>10 antigens) in a single low cost POC device through an innovative disposable and scalable architecture, based on flat panel display technology. Here, we demonstrate multiplexed detection of antibodies to the HPV16 proteins E2, E6, and E7, which are circulating biomarkers for cervical as well as head and neck cancers. This detection technology has 100 percent correlation to our current laboratory-based measurement instrumentation. AUTHORS' DISCLOSURES OF POTENTIAL CONFLICTS OF INTEREST: Benjamin A. Katchman Patents, Royalties, Other Intellectual Property: Arizona State University Joseph T. Smith Patents, Royalties, Other Intellectual Property: Arizona State University Jennifer Blain Christen Patents, Royalties, Other Intellectual Property: Arizona State University Karen S. Anderson Stock or Other Ownership: Provista Diagnostics Consulting or Advisory Role: Provista Diagnostics Patents, Royalties, Other Intellectual Property: Arizona State University


The Analyst ◽  
2016 ◽  
Vol 141 (12) ◽  
pp. 3898-3903 ◽  
Author(s):  
Maowei Dou ◽  
Juan Lopez ◽  
Misael Rios ◽  
Oscar Garcia ◽  
Chuan Xiao ◽  
...  

A low-cost b̲a̲ttery-powered s̲pectrophotometric s̲ystem (BASS) was developed for high-sensitivity point-of-care analysis in low-resource settings on a microfluidic chip without relying on external power supplies.


Sensors ◽  
2018 ◽  
Vol 19 (1) ◽  
pp. 77 ◽  
Author(s):  
Qiao Yu ◽  
Fenfen Zhai ◽  
Hong Zhou ◽  
Zonghua Wang

Basing on the conformation change of aptamer caused by proteins, a simple and sensitive protein fluorescent assay strategy is proposed, which is assisted by the isothermal amplification reaction of polymerase and nicking endonuclease. In the presence of platelet-derived growth factor (PDGF-BB), the natural conformation of a DNA aptamer would change into a Y-shaped complex, which could hybridize with a molecular beacon (MB) and form a DNA duplex, leading to the open state of the MB and generating a fluorescence signal. Subsequently, with further assistance of isothermal recycling amplification strategies, the designed aptamer sensing platform showed an increment of fluorescence. As a benefit of this amplified strategy, the limit of detection (LOD) was lowered to 0.74 ng/mL, which is much lower than previous reports. This strategy not only offers a new simple, specific, and efficient platform to quantify the target protein in low concentrations, but also shows a powerful approach without multiple washing steps, as well as a precious implementation that has the potential to be integrated into portable, low-cost, and simplified devices for diagnostic applications.


2020 ◽  
Vol 187 (9) ◽  
Author(s):  
Bergoi Ibarlucea ◽  
Arnau Pérez Roig ◽  
Dmitry Belyaev ◽  
Larysa Baraban ◽  
Gianaurelio Cuniberti

Abstract A flexible sensor is presented for electrochemical detection of ascorbic acid in sweat based on single-step modified gold microelectrodes. The modification consists of electrodeposition of alginate membrane with trapped CuO nanoparticles. The electrodes are fabricated at a thin polyimide support and the soft nature of the membrane can withstand mechanical stress beyond requirements for skin monitoring. After characterization of the membrane via optical and scanning electron microscopy and cyclic voltammetry, the oxidative properties of CuO are exploited toward ascorbic acid for amperometric measurement at micromolar levels in neutral buffer and acidic artificial sweat, at ultralow applied potential (− 5 mV vs. Au pseudo-reference electrode). Alternatively, measurement of the horizontal shift of redox peaks by cyclic voltammetry is also possible. Obtaining a limit of detection of 1.97 μM, sensitivity of 0.103 V log (μM)−1 of peak shift, and linear range of 10–150 μM, the effect of possible interfering species present in sweat is minimized, with no observable cross-reaction, thus maintaining a high degree of selectivity despite the absence of enzymes in the fabrication scheme. With a lateral flow approach for sample delivery, repeated measurements show recovery in few seconds, with relative standard deviation of about 20%, which can serve to detect increased loss or absence of vitamin, and yet be improved in future by optimized device designs. This sensor is envisioned as a promising component of wearable devices for e.g. non-invasive monitoring of micronutrient loss through sweat, comprising features of light weight, low cost, and easy fabrication needed for such application.


2016 ◽  
Vol 83 (4) ◽  
Author(s):  
Lars D. Renner ◽  
Jindong Zan ◽  
Linda I. Hu ◽  
Manuel Martinez ◽  
Pedro J. Resto ◽  
...  

ABSTRACT An estimated 1.5 billion microbial infections occur globally each year and result in ∼4.6 million deaths. A technology gap associated with commercially available diagnostic tests in remote and underdeveloped regions prevents timely pathogen identification for effective antibiotic chemotherapies for infected patients. The result is a trial-and-error approach that is limited in effectiveness, increases risk for patients while contributing to antimicrobial drug resistance, and reduces the lifetime of antibiotics. This paper addresses this important diagnostic technology gap by describing a low-cost, portable, rapid, and easy-to-use microfluidic cartridge-based system for detecting the ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.) bacterial pathogens that are most commonly associated with antibiotic resistance. The point-of-care molecular diagnostic system consists of a vacuum-degassed microfluidic cartridge preloaded with lyophilized recombinase polymerase amplification (RPA) assays and a small portable battery-powered electronic incubator/reader. The isothermal RPA assays detect the targeted ESKAPE pathogens with high sensitivity (e.g., a limit of detection of ∼10 nucleic acid molecules) that is comparable to that of current PCR-based assays, and they offer advantages in power consumption, engineering, and robustness, which are three critical elements required for the point-of-care setting. IMPORTANCE This paper describes a portable system for rapidly identifying bacteria in resource-limited environments; we highlight the capabilities of the technology by detecting different pathogens within the ESKAPE collection, which cause nosocomial infections. The system is designed around isothermal DNA-based assays housed within an autonomous plastic cartridge that are designed with the end user in mind, who may have limited technological training. Displaying excellent sensitivity and specificity, the assay systems that we demonstrate may enable future diagnoses of bacterial infection to guide the development of effective chemotherapies and may have a role in areas beyond health where rapid detection is valuable, including in industrial processing and manufacturing, food security, agriculture, and water quality testing.


Cellulose ◽  
2020 ◽  
Vol 27 (13) ◽  
pp. 7691-7701 ◽  
Author(s):  
Anusha Prabhu ◽  
M. S. Giri Nandagopal ◽  
Prakash Peralam Yegneswaran ◽  
Hardik Ramesh Singhal ◽  
Naresh Kumar Mani

Abstract We present a high resolution, ultra-frugal printing of paper microfluidic devices using in-house paraffin formulation on a simple filter paper. The patterns printed using an office inkjet printer formed a selective hydrophobic barrier of 4 ± 1 µm thickness with a hydrophilic channel width of 275 µm. These printed patterns effectively confine common aqueous solutions and solvents, which was verified by solvent compatibility studies. SEM analysis reveals that the solvent confinement is due to pore blockage in the filter paper. The fabricated paper-based device was validated for qualitative assessment of Candida albicans (pathogenic fungi) by using a combination of L-proline β-naphthylamide as the substrate and cinnamaldehyde as an indicator. Our studies reveal that the pathogenic fungi can be detected within 10 min with the limit of detection (LOD) of 0.86 × 106 cfu/mL. Owing to its simplicity, this facile method shows high potential and can be scaled up for developing robust paper-based devices for biomarker detection in resource-limited settings. Graphic abstract


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