Enhancement of the Detection Performance of Paper-Based Analytical Devices by Nanomaterials

Paper-based analytical devices (PADs), including lateral flow assays (LFAs), dipstick assays and microfluidic PADs (μPADs), have a great impact on the healthcare realm and environmental monitoring. This is especially evident in developing countries because PADs-based point-of-care testing (POCT) enables to rapidly determine various (bio)chemical analytes in a miniaturized, cost-effective and user-friendly manner. Low sensitivity and poor specificity are the main bottlenecks associated with PADs, which limit the entry of PADs into the real-life applications. The application of nanomaterials in PADs is showing great improvement in their detection performance in terms of sensitivity, selectivity and accuracy since the nanomaterials have unique physicochemical properties. In this review, the research progress on the nanomaterial-based PADs is summarized by highlighting representative recent publications. We mainly focus on the detection principles, the sensing mechanisms of how they work and applications in disease diagnosis, environmental monitoring and food safety management. In addition, the limitations and challenges associated with the development of nanomaterial-based PADs are discussed, and further directions in this research field are proposed.     

Paper-based electrochemiluminescent 3D immunodevice for lab-on-paper, specific, and sensitive point-of-care testing

Recent research on microfluidic paper-based analytical devices (μPADs) has shown that paper has great potential for the fabrication of low-cost diagnostic devices for healthcare and environmental monitoring applications. Herein, electrochemiluminescence (ECL) was introduced for the first time into μPADs that were based on screen-printed paper-electrodes. To further perform high-specificity, high-performance, and high-sensitivity ECL on μPADs for point-of-care testing (POCT), ECL immunoassay capabilities were introduced into a wax-patterned 3D paper-based ECL device, which was characterized by SEM, contact-angle measurement, and electrochemical impedance spectroscopy. With the aid of a home-made device-holder, the ECL reaction was triggered at room temperature. By using a typical tris(bipyridine)ruthenium-tri-n-propylamine ECL system, this paper-based ECL 3D immunodevice was applied to the diagnosis of carcinoembryonic antigens in real clinical serum samples. This contribution further expands the number of sensitive and specific detection modes of μPADs.     

Distance versus Capillary Flow Dynamics-Based Detection Methods on a Microfluidic Paper-Based Analytical Device (μPAD)

In recent years, there has been high interest in paper-based microfluidic sensors or microfluidic paper-based analytical devices (μPADs) towards low-cost, portable, and easy-to-use sensing for chemical and biological targets. μPAD allows spontaneous liquid flow without any external or internal pumping, as well as an innate filtration capability. Although both optical (colorimetric and fluorescent) and electrochemical detection have been demonstrated on μPADs, several limitations still remain, such as the need for additional equipment, vulnerability to ambient lighting perturbation, and inferior sensitivity. Herein, alternative detection methods on μPADs are reviewed to resolve these issues, including relatively well studied distance-based measurements and the newer capillary flow dynamics-based method. Detection principles, assay performance, strengths, and weaknesses are explained for these methods, along with their potential future applications towards point-of-care medical diagnostics and other field-based applications.     

Lab-on-a-chip based immunosensor principles and technologies for the detection of cardiac biomarkers: a review

This review examines the current state of the art lab-on-a-chip and microfluidic based biosensor technologies used in the detection of cardiac biomarkers. The determination and quantification of blood based, cardiac biomarkers are crucial in the triage and management of a range of cardiac related conditions, where time delay has a major impact on short and longer-term outcomes of a patient. The design and manufacturing of biomarker detection systems are multi-disciplinary in nature and require researchers to have knowledge of both life sciences and engineering for the full potential of this field to be realised. This review will therefore provide a comprehensive overview of chip based immunosensing technology as applied to cardiac biomarker detection, while discussing the potential suitability and limitations of each configuration for incorporation within a clinical diagnostics device suitable for point-of-care applications.     

Simple and covalent fabrication of a paper device and its application in sensitive chemiluminescence immunoassay

In this paper, chemiluminescence immunoassay (CLIA) was introduced into the recently proposed microfluidic paper-based analytical devices (μPADs) through covalent fabrication strategy for the first time. This novel paper-based CLIA, with high-throughput, rapid, stable and reusable CL response to trace amounts of analyte in real biological samples, combines the simplicity and low-cost of the μPADs with the high sensitivity and selectivity of CLIA. Periodate oxidation, which can form covalent bonds between polysaccharides and proteins, was used for activation of μPADs to covalently immobilize antibodies on μPADs in this work for the first time. Thus, the paper-based sandwich CLIA and regeneration of it can be easily realized for further development of this technique in sensitive, specific and low-cost applications. The application test of this paper-based CLIA was successfully performed, as a model, through the determination of biomarkers in human serum on paper microzone plate. The paper-based CLIA will be very useful when the level of an analyte in real biological sample is important for point-of-care testing, public health and environmental monitoring in remote regions, developing or developed countries.     

Electrochemical Immunosensors: The Evolution from Elisa to EμPADs

Electrochemical immunosensors comprise the merging of two different disciplines: molecular biology and electrochemistry. This review explains in depth the main parts of electrochemical immunosensors and how the enzyme-linked immunosorbent assay (ELISA) has been integrated into sophisticated “lab-on-a-chip” and “point-of-care” devices. It also reviews how nanotechnology has been a powerful tool for achieving lower detection limits, more signal amplification, and constructing label-free devices. It finally explores the new perspectives on electrochemical immunosensors to integrate them in novel paper microfluidic devices called EμPADs. Colleagues introducing themselves to the topic for the first time will find in this review a comprehensive revision of how the basics of the technology have given rise to the emerging topic of EμPADs.     

Silicon nanowire biosensor and its applications in disease diagnostics: A review

Over the past decade, silicon nanowire (SiNW) biosensors have been studied for the detection of biological molecules as highly sensitive, label-free, and electrical tools. Herein we present a comprehensive review about the fabrication of SiNW biosensors and their applications in disease diagnostics. We discuss the detection of important biomarkers related to diseases including cancer, cardiovascular diseases, and infectious diseases. SiNW biosensors hold great promise to realize point-of-care (POC) devices for disease diagnostics with potential for miniaturization and integration.     

Miniaturized surface engineered technologies for multiplex biosensing devices

The demand for quick, accurate, and affordable point-of-care (POC) devices increases with the advancement in the dimensions of nanotechnology and digital interfaces (Internet of Things). The future of diagnostic requires the platform which can provide us the following benefits i. e., on-site detection, qualitative as well as quantitative analysis, easy to use, portable, low sample requirement, cost-effective, and have multiplexing proficiency. Multiplex biosensing platforms (MBPs) have the above following advantages so are going to be mostly used in various healthcare applications in near future. MBPs have the potential to fulfill the ‘ASSURED’ criteria specified by the World Health Organization (WHO) for remote-limited settings. This review paper focuses on miniaturized platforms that have multiplexing benefits for the bioanalysis of different clinical samples related to various healthcare applications. In addition to this, screening of pesticides, antibiotics, and hazardous metal ions with these surface-engineered devices has also been accounted in food and environmental samples. Some of the advanced techniques including microfluidics (Lab-on-a-chip), wearable smart devices, and CRISPR/Cas system for multiplexing applications are briefly described here. Furthermore, various needs, challenges, and prospects in commercializing these multiplexed surface-engineered devices have been discussed in this review.     

普适型纸芯片的研究与应用

纸基分析芯片(纸芯片)具有成本低、便携化、操作和后处理简单无污染等优点,在临床诊断、食品质量控制和环境监测等领域有着广阔的应用前景。然而,由于难以制作性能优异的疏脂性屏障,使得纸芯片在涉及有机溶剂和表面活性剂的分析检测中,其发展受到了限制。针对当前纸芯片开发研究中存在的灵敏度较低、对有机溶剂和表面活性剂敏感等难点问题,研究人员在滤纸基底上制作出了性能优异的疏水隔离图案和高粘附疏水表面,并验证了所制备的纸芯片对涉及有机溶剂和表面活性剂的分析检测具有普适性。本文对此类普适型纸芯片的研究与应用进行评述。     

Microfluidic systems for the analysis of blood-derived molecular biomarkers

Biomarkers are relevant indicators of the physiological state of an individual. Although biomarkers can be found in diseased tissue and different biofluids, sampling from blood plasma is relatively easy and less invasive. Among the molecular biomarkers that can be found circulating in plasma are proteins, metabolites, nucleic acids, and exosomes. Some of these plasma-circulating biomarkers are now employed for patient stratification in a broad range of diseases with high sensitivity and specificity and are useful in early diagnosis, initial risk assessment, and therapy selection. However, there is a pressing need to develop novel approaches for biomarker analysis that can be translated into clinical or other settings without complex methodologies or instrumentation. Microfluidics has been touted as a promising technology to carry out this task because it offers high-throughput, automation, multiplexed detection, and portability, possibly overcoming the bottleneck that prevent the translation of novel biomarkers to the point-of-care (POC). Here, we provide a review of the microfluidic systems that have been engineered to detect circulating molecular biomarkers in blood plasma. We also review the different microfluidic approaches for plasma enrichment, which are now being integrated with microfluidic-based biomarker analyzers. Such integration should lead to cost-effective solutions in in vitro diagnostics, with special relevance to POC platforms.     

Cancer detection using nanoparticle-based sensors

This tutorial review surveys the latest achievements in the use of nanoparticles to detect cancer biomarkers and cancer cells with a focus on optical and electrochemical techniques. Nanoparticle based cancer diagnostics are becoming an increasingly relevant alternative to traditional techniques. Although some drawbacks exist in relation to the obtained sensitivity the use of nanoparticle-based sensors in biomarker detection or cancer cell detection offers some advantages in comparison to conventional methods. The developed techniques can be interesting and relevant for their use in point-of-care of cancer diagnostics. The methods can be of low cost and in addition easy to be incorporated into user-friendly sensing platforms.     

Paper‐Based Electrochemiluminescent 3D Immunodevice for Lab‐on‐Paper,Specific, and Sensitive Point‐of‐Care Testing

Recent research on microfluidic paper‐based analytical devices (μPADs) has shown that paper has great potential for the fabrication of low‐cost diagnostic devices for healthcare and environmental monitoring applications. Herein, electrochemiluminescence (ECL) was introduced for the first time into μPADs that were based on screen‐printed paper‐electrodes. To further perform high‐specificity, high‐performance, and high‐sensitivity ECL on μPADs for point‐of‐care testing (POCT), ECL immunoassay capabilities were introduced into a wax‐patterned 3D paper‐based ECL device, which was characterized by SEM, contact‐angle measurement, and electrochemical impedance spectroscopy. With the aid of a home‐made device‐holder, the ECL reaction was triggered at room temperature. By using a typical tris(bipyridine)ruthenium–tri‐n‐propylamine ECL system, this paper‐based ECL 3D immunodevice was applied to the diagnosis of carcinoembryonic antigens in real clinical serum samples. This contribution further expands the number of sensitive and specific detection modes of μPADs.     

Cellphone-based devices for bioanalytical sciences

During the last decade, there has been a rapidly growing trend toward the use of cellphone-based devices (CBDs) in bioanalytical sciences. For example, they have been used for digital microscopy, cytometry, read-out of immunoassays and lateral flow tests, electrochemical and surface plasmon resonance based bio-sensing, colorimetric detection and healthcare monitoring, among others. Cellphone can be considered as one of the most prospective devices for the development of next-generation point-of-care (POC) diagnostics platforms, enabling mobile healthcare delivery and personalized medicine. With more than 6.5 billion cellphone subscribers worldwide and approximately 1.6 billion new devices being sold each year, cellphone technology is also creating new business and research opportunities. Many cellphone-based devices, such as those targeted for diabetic management, weight management, monitoring of blood pressure and pulse rate, have already become commercially-available in recent years. In addition to such monitoring platforms, several other CBDs are also being introduced, targeting e.g., microscopic imaging and sensing applications for medical diagnostics using novel computational algorithms and components already embedded on cellphones. This report aims to review these recent developments in CBDs for bioanalytical sciences along with some of the challenges involved and the future opportunities.

Microfluidic designs and techniques using lab-on-a-chip devices for pathogen detection for point-of-care diagnostics

Effective pathogen detection is an essential prerequisite for the prevention and treatment of infectious diseases. Despite recent advances in biosensors, infectious diseases remain a major cause of illnesses and mortality throughout the world. For instance in developing countries, infectious diseases account for over half of the mortality rate. Pathogen detection platforms provide a fundamental tool in different fields including clinical diagnostics, pathology, drug discovery, clinical research, disease outbreaks, and food safety. Microfluidic lab-on-a-chip (LOC) devices offer many advantages for pathogen detection such as miniaturization, small sample volume, portability, rapid detection time and point-of-care diagnosis. This review paper outlines recent microfluidic based devices and LOC design strategies for pathogen detection with the main focus on the integration of different techniques that led to the development of sample-to-result devices. Several examples of recently developed devices are presented along with respective advantages and limitations of each design. Progresses made in biomarkers, sample preparation, amplification and fluid handling techniques using microfluidic platforms are also covered and strategies for multiplexing and high-throughput analysis, as well as point-of-care diagnosis, are discussed.     

USB powered microfluidic paper-based analytical devices

Microfluidic paper-based analytical devices (μPADs) allow user-friendly and portable chemical determinations, although they provide limited applicability due to insufficient sensitivity. Several approaches have been proposed to address poor sensitivity in μPADs, but they frequently require bulky equipment for power and/or read-outs. Universal serial buses (USB) are an attractive alternative to less portable power sources and are currently available in many common electronic devices. Here, USB-powered μPADs (USB μPADs) are proposed as a fusion of both technologies to improve performance without adding instrumental complexity. Two ITP USB μPADs were developed, both powered by a 5 V potential provided through standard USB ports. The first device was fabricated using the origami approach. Its operation was analyzed experimentally and numerically, yielding a two-order-of-magnitude sample focusing in 15 min. The second ITP USB μPAD is a novel design, which was numerically prototyped with the aim of handling larger sample volumes. The reservoirs were moved away from the ITP channel and capillary action was used to drive the sample and electrolytes to the separation zone, predicting 25-fold sample focusing in 10 min. USB μPADs are expected to be adopted by minimally-trained personnel in sensitive areas like resource-limited settings, the point-of-care and in emergencies.     

Nanomaterial Based Biosensors for Detection of Biomarkers of Exposure to OP Pesticides and Nerve Agents: A Review

Organophosphorus (OP) pesticides are primarily used as insecticides and chemical warfare agents worldwide. Due to their impact on the environment and health, it is important to develop prompt and accurate pesticide analysis method. This review addresses recent advances and new trends in nanotechnology‐based biosensors for biological monitoring of exposures to OP pesticides and nerve agents. In order to determine them, we have to find the corresponding biomarkers. In 1989, the national academy of sciences (NAS)divided biomarkers into the following three categories: biomarker of exposure, biomarker of effect and biomarker of susceptibility (Figure 1A). The unique chemical and physical properties of nanomaterial have paved the way to new and improved sensing devices, in general, and electrochemical/optical biosensors, in particular. In this paper, background information and a general overview of electrochemical/immunoassay detection techniques are provided. Various nanomaterial labels are discussed. Usually nanomaterials can be roughly divided into nanometer powder, nanometer fiber, nanometer film, nanometer block and so on four classes, such as colloidal gold, semiconductor nanoparticles and carbon nanomaterial (Figure 1B). In addition, we discuss some future considerations and opportunities for advancing the use of biosensors for environmental and health studies.     

Integrated dielectrophoretic and impedimetric biosensor provides a template for universal biomarker sensing in clinical samples

The detection and quantification of nucleic acid and proteomic biomarkers in bodily fluids is a critical part of many medical screening and diagnoses. However, majority of the current detection platforms are not ideal for routine, rapid, and low-cost testing in point-of-care settings. To address this issue, we developed a concept for a disposable universal point-of-care biosensor that can detect and quantify nucleic acid and proteomic biomarkers in diluted serum samples. The central tenet of sensing is the use of dielectrophoresis, electrothermal effects, and thermophoresis to selectively and rapidly isolate the biomarkers of interest in electrodes and then quantify using electrical impedance. When the sensor was applied to quantify microRNA and antigen biomarker molecules directly in diluted serum samples, it produced a LOD values in the fM range and sensitivity values from 10¹² to 10¹⁵ Ω/M with a 30 min assay time and assay cost of less than $50 per assay.     

Lab-on-paper-based devices using chemiluminescence and electrogenerated chemiluminescence detection

As an analytical support, paper, being low cost, highly abundant, of high porosity, disposable or biodegradable, and easy to use, store, transport, and print, has excellent chemical compatibility with many applications. Since the first microfluidic paper-based analytical device (μ-PAD or lab-on-paper) was proposed, the paper-based assay has never attracted as much attention as it does now. There has recently been rapidly increasing interest in using sensitive luminescence methods, for example chemiluminescence (CL) and electrogenerated chemiluminescence (ECL), as the detection strategy for lab-on-paper devices. Because of their intrinsic characteristics, CL and ECL provide outstanding performance while retaining the simplicity, low cost, multifunctionality, versatility, flexibility, and disposability of μ-PADs. The objective of this review is to cover the development of lab-on-paper-based devices using CL and ECL detection, including fabrication of paper devices, construction of sensing interfaces, signal amplification strategies, external instruments used, and applications. We believe that lab-on-paper devices with CL and ECL detection methods will meet the diverse requirements of point-of-care diagnosis.     

Enzymatic Methods for Salivary Biomarkers Detection: Overview and Current Challenges

Early detection is a key factor in patient fate. Currently, multiple biomolecules have been recognized as biomarkers. Nevertheless, their identification is only the starting line on the way to their implementation in disease diagnosis. Although blood is the biofluid par excellence for the quantification of biomarkers, its extraction is uncomfortable and painful for many patients. In this sense, there is a gap in which saliva emerges as a non-invasive and valuable source of information, as it contains many of the biomarkers found in blood. Recent technological advances have made it possible to detect and quantify biomarkers in saliva samples. However, there are opportunity areas in terms of cost and complexity, which could be solved using simpler methodologies such as those based on enzymes. Many reviews have focused on presenting the state-of-the-art in identifying biomarkers in saliva samples. However, just a few of them provide critical analysis of technical elements for biomarker quantification in enzymatic methods for large-scale clinical applications. Thus, this review proposes enzymatic assays as a cost-effective alternative to overcome the limitations of current methods for the quantification of biomarkers in saliva, highlighting the technical and operational considerations necessary for sampling, method development, optimization, and validation.