|
BioChip Journal
Journal Prestige (SJR): 0.412  Citation Impact (citeScore): 1 Number of Followers: 0  Hybrid journal (It can contain Open Access articles)ISSN (Print) 1976-0280 - ISSN (Online) 2092-7843 Published by Springer-Verlag  [2468 journals]
|
- Advanced Organotypic In Vitro Model Systems for Host–Microbial
Coculture-
Free pre-print version: Loading...
Abstract: In vitro model systems have been advanced to recapitulate important physiological features of the target organ in vivo more closely than the conventional cell line cultures on a petri dish. The advanced organotypic model systems can be used as a complementary or alternative tool for various testing and screening. Numerous data from germ-free animal studies and genome sequencings of clinical samples indicate that human microbiota is an essential part of the human body, but current in vitro model systems rarely include them, which can be one of the reasons for the discrepancy in the tissue phenotypes and outcome of therapeutic intervention between in vivo and in vitro tissues. A coculture model system with appropriate microbes and host cells may have great potential to bridge the gap between the in vitro model and the in vivo counterpart. However, successfully integrating two species in one system introduces new variables to consider and poses new challenges to overcome. This review aims to provide perspectives on the important factors that should be considered for developing organotypic bacterial coculture models. Recent advances in various organotypic bacterial coculture models are highlighted. Finally, challenges and opportunities in developing organotypic microbial coculture models are also discussed.
PubDate: 2023-05-22
-
- Paper-Based Radial Flow Assay Integrated to Portable Isothermal
Amplification Chip Platform for Colorimetric Detection of Target DNA-
Free pre-print version: Loading...
Abstract: A novel integrated detection system that introduces a paper-chip-based molecular detection strategy into a polydimethylsiloxane (PDMS) microchip and temperature control system was developed for on-site colorimetric detection of DNA. For the paper chip-based detection strategy, a padlock probe DNA (PLP)-mediated rolling circle amplification (RCA) reaction for signal amplification and a radial flow assay according to the Au-probe labeling strategy for visualization were optimized and applied for DNA detection. In the PDMS chip, the reactions for ligation of target-dependent PLP, RCA, and labeling were performed one-step under isothermal temperature in a single chamber, and one drop of the final reaction solution was loaded onto the paper chip to form a radial colorimetric signal. To create an optimal analysis environment, not only the optimization of molecular reactions for DNA detection but also the chamber shape of the PDMS chip and temperature control system were successfully verified. Our results indicate that a detection limit of 14.7 nM of DNA was achieved, and non-specific DNAs with a single-base mismatch at the target DNA were selectively discriminated. This integrated detection system can be applied not only for single nucleotide polymorphism identification, but also for pathogen gene detection. The adoption of inexpensive paper and PDMS chips allows the fabrication of cost-effective detection systems. Moreover, it is very suitable for operation in various resource-limited locations by adopting a highly portable and user-friendly detection method that minimizes the use of large and expensive equipment.
PubDate: 2023-04-27
-
- Comparison and Analysis of Polymer-Functionalized Carbon Nanotubes for
Enhancement of the Quantitative Detection of Procalcitonin Levels in Human
Plasma-
Free pre-print version: Loading...
Abstract: Procalcitonin (PCT) is one of the core biomarkers for the body’s systemic inflammatory response that is expressed uniformly in several organ tissues. PCT concentration levels rise significantly reaching the highest peak within 48 h after the bacterial infection starts. In this study, we evaluated different polymer-functionalized carbon nanotubes (CNTs) to enhance the performance of a sensing platform for the rapid detection of PCT over a large physiological range. CNTs were treated with polyethylene glycol (PEG), polyvinylpyrrolidone (PVP), and poly(L-arginine) P(Arg) to improve the nanostructures while promoting stability and reducing non-specific interaction. Different morphology analyses were performed to investigate the properties of the polymer-functionalized CNTs and compared them with the pristine material. Screen-printed carbon electrode (SPCE) sensors were modified with the three proposed nanocomposites and electrical characterization are performed. Afterward, the PCT antibody (100 ng/ml) is immobilized on the working electrodes before the electrochemical measurements with the analytes. Differential pulse voltammetry (DPV) responses were measured to quantify the PCT (27.43–20,000 pg/ml) in buffer solution and human plasma samples. CNT-PVP and CNT-P(Arg) electrodes presented a high correlation between the current peaks and the concentration of PCT. Subsequently, an interference study was carried out with inflammatory-related biomarkers that could cause a false positive response, the variation of the signal current was less than 3%. The SPCE devices also went through validation with the “gold-standard” laboratory method to verify the agreement with the proposed method. This study regarding the optimization of the CNTs is just the first part of our final target regarding a multiplexed point-of-care device using a small volume of the blood sample to provide a rapid diagnostic regarding inflammatory response.
PubDate: 2023-04-27
-
- A Laser-Micromachined PCB Electrolytic Micropump Using an Oil-Based
Electrolyte Separation Barrier-
Free pre-print version: Loading...
Abstract: We report a laser-micromachined electrolytic PCB micropump with an oil separation barrier. As advances in terms of miniaturization and performance from our previous mesoscale PCB electrolytic pump (Kim et al. in Sens Actuators A Phys 277:73–84, 2018), we employed a simple yet rapid tape-based laser-machining technique called tape-liner-supported plastic laser micromachining and pattern transfer to fabricate a microfluidic coverslip for a PCB electrode chip. Using our microfabrication technique, the coverslip is bonded to a PCB chip to form an enclosed microscale pump with a high machining precision and no need for alignment of intermediate adhesive tapes with structural layers as commonly done in previous tape-bonding work. The completed micropump demonstrated excellent pumping performance: flow rate up to 24.49 ml/min and backpressure up to 394 kPa. Electrochemical activation of electrodes consisting of a train of voltage pulses and sweeps improves the pumping performance. In order to prevent unwanted interspersion between the electrolyte and working fluid, various separation diaphragms were previously employed, but at the cost of limited working volume and flow rate as the diaphragms were permanently anchored to the pump body. Here we propose to use an oil plug as an untethered (mobile) separation barrier. After a systematic study of properties of common oils, we tested fluorinated oil (HFE-7500), hexadecane, and tetradecane as the candidate barrier materials. HFE-7500 was chosen because its interface was stable and did not degrade pumping performance for the flow-rate range of 8.47 μl/min–2.48 ml/min. We expect our micropump with the oil plug to be used as an excellent pressure source for integrated lab-on-a-chip devices, especially lab-on-a-PCBs.
PubDate: 2023-03-30
-
- Fabrication of Cell Spheroids for 3D Cell Culture and Biomedical
Applications-
Free pre-print version: Loading...
Abstract: The significance of 3D cell culture—i.e., the maintenance of cell characteristics by mimicking the natural environment—has been increasing, and various 3D cell culture methods have been proposed. 3D cell culture is a widely used cell spheroid culture method, which is a representative method of matrix-free 3D cell culture. In contrast with that in conventional 2D cell culture, cells aggregate in a spherical shape with abundant cell–cell and cell–matrix interactions that more closely resemble the in vivo environment in a 3D culture. In a cell spheroid culture, the viability and functionality of spheroids change based on the cell type and morphology, and their controllable characteristics differ depending on the fabrication method used. Moreover, the mass production and size controllability of spheroids are dependent on the fabrication method used; therefore, the spheroid utilization method is also different. In this review, representative cell spheroid fabrication methods are examined, and the possibility of standardization is discussed so that these methods can be widely used. In addition, the biomedical applications of spheroids and their potential for future development are explored.
PubDate: 2023-03-01
-
- Development of an Integrated Biochip System Consisting of a Magnetic
Particle Washing Station and a Markerless Volumetric Biochip-
Free pre-print version: Loading...
Abstract: As the use of markers in conventional volumetric biochips results in complex biochip structures and assay processes, an integrated biochip system consisting of a magnetic particle (MP) washing station and a markerless volumetric biochip was developed in this study. The biochip was composed of a reaction chamber and a channel connected to the upper side of the chamber. The assay process involved the injection of a sample containing a catalase-labeled analyte into the reaction chamber containing a substrate solution comprising H2O2, detergent, and methylene blue; and the measurement of the distance traveled by the generated foam. The MP washing station had two check valves installed at different ports of a T-connector in opposite directions. Contaminants were removed from the MPs by pressing and releasing the pipette plunger, which caused a one-way flow of washing buffer from the buffer chamber to the waste chamber through the pipette. The integrated system was applied for the detection of Salmonella typhimurium using magnetic capture particles (CPs) functionalized with anti-S. typhimurium antibodies and labeling particles (LPs) functionalized with catalase and antibodies. After the bacterial sample was mixed with CPs and LPs, the resulting CP–bacteria–LP complex was washed in the MP washing station and analyzed in the markerless volumetric biochip. This method enabled the detection of bacteria with a limit of detection of 1.6 CFU within 25 min. Considering the high sensitivity and simplicity of the biochip structure and the assay process, the integrated system demonstrates considerable potential for the point-of-care testing of various analytes.
PubDate: 2023-03-01
-
- Recent Advances in Electrochemical and Optical Biosensors for Cancer
Biomarker Detection-
Free pre-print version: Loading...
Abstract: Cancer is a universal disease with a high mortality rate and caused by an uncontrolled growth of abnormal cells. Each cancer has its unique molecular change such as up/down-regulations of biological molecules in cancer cells. These biological molecules have been identified as biomarkers and used as a target analyte for cancer diagnosis. Since the level of cancer biomarkers is very insufficient at an early stage cancer, there is a need for technological advances that can be more accurate, ultra-sensitive, trustworthy, and selective to biomarkers. This review summarizes evaluation methods and recent advances in electrochemical and optical (fluorescent and colorimetric) biosensors to detect various cancer biomarkers as molecular targets. Electrochemical methods demonstrate rapid and ultra-sensitive detection of cancer biomarkers. And, fluorescent biosensors present improved target detection sensitivity by signal amplification strategy as well as provide the possibility to sense multiple biomarker targets with multiple fluorescent probes. Colorimetric biosensors easily and quickly detect cancer biomarkers by the naked eye for use as a point-of-care testing (POCT) platform. These biosensors can be further improved by understanding the complication of cancer cells and the molecular alterations in cancer progression for early diagnosis of cancer and patient prognosis.
PubDate: 2023-03-01
-
- Organ-On-A-Chip Technology: An In-depth Review of Recent Advancements and
Future of Whole Body-on-chip-
Free pre-print version: Loading...
Abstract: One of the leading challenges facing the pharmaceutical industry today is the ‘high attrition rate problem’, with a large number of drug candidates being abandoned before reaching the clinical stage. The high failure rate is not only costly for the pharmaceutical industry but also hinders the access of patients to better treatment options. This ever-widening gap can be attributed to the current in-vitro and animal testing techniques for predicting human responses to drug candidates. Two-dimensional (2D) in-vitro cell culture fails to recapitulate key physical and biochemical cues while phylogenetic differences between animal models and humans in key physiological systems make data generated by these platforms unreliable. There have been several solutions proposed but one that has been gaining a lot of prominence are microfluidic-based microsystems able to mimic organs, called ‘Organ-On-A-Chip’ devices and is built upon the principles of microfluidics, material science, and cell biology. Here we delve into the ongoing research involving this platform demonstrating its key strengths and the application of this technology in several key stages of drug discovery such as target identification and high throughput screening. We also discuss the potential, future, and key obstacles that lie in the universal adoption of this technology. Graphical
PubDate: 2023-03-01
-
- Development of Tumor-Vasculature Interaction on Chip Mimicking Vessel
Co-Option of Glioblastoma-
Free pre-print version: Loading...
Abstract: Vessel co-option (VC) differs from angiogenesis in that tumor cells grow toward blood vessels. Through VC, tumor cells can receive relatively more nutrients and oxygen from blood vessels. Despite its clinical significance, VC is relatively less studied compared to angiogenesis because of difficulties in longitudinal observation of VC in vivo and lack of proper VC models in vitro. A needle template method in which microchannels are formed in hydrogel by needles was used to form blood vessels and mimic angiogenesis. However, it has not yet been used to mimic VC. In this study, we report the development of VC on chip based on the needle template method. On the VC on chip, the effect of distance between spheroids and blood vessels on VC induction was investigated by seeding glioblastoma (GBM) spheroids 50 and 250 μm from the preformed blood vessels. Irrespective of distance, cancer cells from the spheroids grew toward the blood vessels but did not penetrate the vessels, indicating that GBM cells showed VC-like behavior. These results suggest that our chip could recapitulate VC in GBM.
PubDate: 2023-03-01
-
- Development of Gut-Mucus Chip for Intestinal Absorption Study
-
Free pre-print version: Loading...
Abstract: The intestinal epithelium is a major barrier through which orally administered drugs must pass. The intestinal mucosa on the epithelium acts as an additional barrier that protects the intestinal cells from foreign substances, thereby interfering with drug delivery. Caco-2 based cell culture model is a standard in vitro model system for testing drug uptake. However, current in vitro models do not reflect the absorption mechanism of drugs accurately due to the absence of the mucus layer. Here, we developed a microfluidic gut-mucus chip using Caco-2 cells coated with mucin protein. It was confirmed that the mucin layer was maintained under flow conditions by Alcian blue/Periodic Acid Schiff (PAS) staining. In addition, the effect of mucosal layer on drug absorption in the flow environment was examined. Mucus-adhesive particles can be useful for delivery of drugs across the intestinal epithelium. We prepared mucus-adhesive and non-adhesive microparticles containing fluorescent molecules and compared the adhesion of these particles in flow condition. Mucus-coated Caco-2 cells provide a more physiologically realistic intestinal epithelial environment to study uptake processes of drugs released from the mucus-adhesive particles. We hope that the gut-mucus chip could potentially be used as novel and more accurate in vitro models of the intestine.
PubDate: 2023-02-15
DOI: 10.1007/s13206-023-00097-0
-
- From Single- to Multi-organ-on-a-Chip System for Studying Metabolic
Diseases-
Free pre-print version: Loading...
Abstract: Metabolic diseases have been a major public health issue and a clinical challenge worldwide. Dysfunction of metabolic homeostasis is linked to the onset of various diseases because the various metabolizing organs synchronously interact to maintain metabolic homeostasis. Therefore, understanding inter-organ communication is important to develop more accurate drug testing and discovery for the treatment of metabolic diseases. The physiologically relevant organ-on-a-chip system is an emerging technology to study metabolic dynamics by providing an organ-level platform. This review summarizes the single- and multi-organ chip systems to study metabolic diseases and their dynamics, focusing on their inter-organ crosstalk.
PubDate: 2023-02-15
DOI: 10.1007/s13206-023-00098-z
-
- Flexible Capillary Microfluidic Devices Based on Surface-Energy Modified
Polydimethylsiloxane and Polymethylmethacrylate with Room-Temperature
Chemical Bonding-
Free pre-print version: Loading...
Abstract: Polydimethylsiloxane (PDMS) has been widely used for the rapid prototyping of microfluidic devices for biosensor cartridges. However, using PDMS to prototype capillary-driven microfluidic devices is often limited by the difficulty of maintaining the surface energy of surface-treated PDMS for an extended period in addition to the degradation of the biosensing elements during the bonding process at elevated temperature. Herein, prototyping of a flexible capillary microfluidic channel (FCMC) device based on the room-temperature bonding of the surface energy-modified PDMS (m-PDMS) microfluidic channel and a thermoplastic lid, polymethylmethacrylate (PMMA), is introduced for prolonged control of passive liquid flow characteristics. The m-PDMS was fabricated by blending polydimethylsiloxane-ethylene oxide (60–70%) block copolymer (PDMS-b-PEO) additive with pre-PDMS, of which the water contact angles could be controlled between 38.5° and 78.5° by adjusting the ratio of the two components. Room-temperature bonding of the m-PDMS and PMMA sheets functionalized by 3-glycidoxypropyltrimethoxysilane and aminopropyltriethoxysilane, respectively, was introduced to fabricate the FCMC devices via the formation of a stable linker epoxy-amine without the requirement of elevated temperatures. The FCMC device possessed longevity to passively drive liquid in the channel for 2 months under ambient conditions due to the prolonged stable hydrophilicity of m-PDMS. The proposed approaches provide great potential for prototyping passive microfluidic devices for biosensor cartridge applications.
PubDate: 2023-01-31
DOI: 10.1007/s13206-023-00096-1
-
- Deep Learning-Assisted Droplet Digital PCR for Quantitative Detection of
Human Coronavirus-
Free pre-print version: Loading...
Abstract: Since coronavirus disease 2019 (COVID-19) pandemic rapidly spread worldwide, there is an urgent demand for accurate and suitable nucleic acid detection technology. Although the conventional threshold-based algorithms have been used for processing images of droplet digital polymerase chain reaction (ddPCR), there are still challenges from noise and irregular size of droplets. Here, we present a combined method of the mask region convolutional neural network (Mask R-CNN)-based image detection algorithm and Gaussian mixture model (GMM)-based thresholding algorithm. This novel approach significantly reduces false detection rate and achieves highly accurate prediction model in a ddPCR image processing. We demonstrated that how deep learning improved the overall performance in a ddPCR image processing. Therefore, our study could be a promising method in nucleic acid detection technology.
PubDate: 2023-01-17
DOI: 10.1007/s13206-023-00095-2
-
- Identification of Potential Allergens of Atractylodes japonica and
Addition of Panels for Allergic Diseases-
Free pre-print version: Loading...
Abstract: The use of herbal medicines is increasing significantly worldwide. However, studies on side effects such as allergic reactions of herbal medicines are lacking. To provide an effective prescription, a diagnostic test to determine whether an allergic reaction is induced in the patient must be preceded. In this study, the purpose of this study is to investigate whether Atractylodes japonica extract, used as herbal medicine, binds to human serum IgE and induces an allergic reaction as an immune response complex, and to add a panel of allergens to the Allergy Q kit. First, using the Allergy-Q test kit, 253 sera samples were screened and 4 samples each from positive and negative serogroups were randomly selected. AJ extracts were separated by size through sodium dodecyl sulfate–polyacrylamide gel electrophoresis and stained with Coomassie blue dye. We then used Western blotting and in-gel digest to identify proteins that bind to human serum IgE, and predicted six proteins, including putative chitinases, proteasomes, and hypothetical proteins, using MASCOT program matching. Consequently, we demonstrated the potential of AJ as an allergen by confirming the sensitivity between AJ-derived proteins and human IgE.
PubDate: 2023-01-17
DOI: 10.1007/s13206-022-00094-9
-
- Fluorescent Lateral Flow Assay with Carbon Nanodot Conjugates for
Carcinoembryonic Antigen-
Free pre-print version: Loading...
Abstract: A lateral flow assay (LFA) sensor on a half-strip platform labeled with blue carbon nanodot-polystyrene (PS-CND) nanoconjugates was developed for the detection of carcinoembryonic antigen (CEA) levels in buffer and serum solutions from healthy and cancer patients. CNDs, biocompatible nanoparticles containing amino groups synthesized by hydrothermal synthesis, were conjugated to spherical polystyrene (PS) beads with an average diameter of 60 nm, followed by the attachment of a detection probe, anti-CEA (M0911042), using a heterobifunctional cross-linker. PS beads were used as a template in CND conjugates to provide uniform size and shape of fluorescent labels without losing CND fluorescence intensity after the antibody conjugation step and to improve fluorescence stability. Upon the interaction of CEA from samples with the anti-CEA (M0911042) probe-modified PS-CND, which was further adsorbed onto a test line composed of the capture anti-CEA (M0911041) physisorbed onto a nitrocellulose membrane, the fluorescent signals on the test line increased as a function of the CEA concentrations under irradiation with a portable 365 nm ultraviolet lamp. A linear concentration range of 0.04–70 nM in buffer was observed, with a limit of detection of 0.3 nM. The applicability of the developed LFA half-strip sensor for disease diagnosis was demonstrated by identifying fluorescent levels on the test line due to the presence of CEA in serum samples from cancer patients. Importantly, signals from healthy human serum solutions because of lower CEA concentrations beyond the sensor detection capability were clearly distinguished from the patient ones.
PubDate: 2023-01-17
DOI: 10.1007/s13206-022-00093-w
-
- Polydopamine Nanoparticle-Incorporated Fluorescent Hydrogel for
Fluorescence Imaging-Guided Photothermal Therapy of Cancers-
Free pre-print version: Loading...
Abstract: In this study, we report a fluorescent polydopamine nanoparticle-incorporated hydrogel (FPNP-hydrogel) for fluorescence imaging-guided photothermal cancer therapy. The FPNP-hydrogel was prepared by combining fluorescent polydopamine nanoparticles (FPNPs) with polyvinyl alcohol (PVA) using a simple method. The unique optical properties of the FPNPs facilitated the FPNP-hydrogel the excellent biocompatibility, strong fluorescence, and high photostability, showing an availability of fluorescence imaging in BALB/c nude mice. When combined with near-infrared laser irradiation, the FPNP-hydrogel noticeably reduced tumor volumes in 4T1 tumor-bearing mice within 14 days without inducing the inflammatory response in major organs. In conclusion, the results suggest that multifunctional hydrogels with excellent fluorescence properties and high photothermal conversion efficiency could be useful for designing novel therapeutic methods for cancer treatment.
PubDate: 2022-12-06
DOI: 10.1007/s13206-022-00091-y
-
- Single-Shot Light-Field Microscopy: An Emerging Tool for 3D Biomedical
Imaging-
Free pre-print version: Loading...
Abstract: 3D microscopy is a useful tool to visualize the detailed structures and mechanisms of biomedical specimens. In particular, biophysical phenomena such as neural activity require fast 3D volumetric imaging because fluorescence signals degrade quickly. A light-field microscope (LFM) has recently attracted attention as a high-speed volumetric imaging technique by recording 3D information in a single-snapshot. This review highlighted recent progress in LFM techniques for 3D biomedical applications. In detail, various image reconstruction algorithms according to LFM configurations are explained, and several biomedical applications such as neuron activity localization, live-cell imaging, locomotion analysis, and single-molecule visualization are introduced. We also discuss deep learning-based LFMs to enhance image resolution and reduce reconstruction artifacts.
PubDate: 2022-12-01
DOI: 10.1007/s13206-022-00077-w
-
- Microfabricated Stretching Devices for Studying the Effects of Tensile
Stress on Cells and Tissues-
Free pre-print version: Loading...
Abstract: Tensile stress is one of the most common mechanical stresses on the connective tissues of human organs. Cell stretching devices have been developed to study the effects of tensile stress on cells and tissues. In this study, we review how these devices function mechanically and apply them to biological research. To this end, we technically evaluate the four types of actuation processes used in cell stretching devices, including electric motor-driven and electromagnetic actuation, along with their pros and cons. For example, these cell stretching devices have shortcomings including large size, a complicated system, and generation of heat and shock, which hinder the real-time imaging of cells during stretching in high-resolution microscopes. We also describe the effects of tensile stress on cellular and tissue homeostasis. With this review, we seek to explore future directions for development of cell tensioning devices to understand mechanobiological responses to mechanical stress in vivo.
PubDate: 2022-12-01
DOI: 10.1007/s13206-022-00073-0
-
- Thermoplasmonic Scaffold Design for the Modulation of Neural Activity in
Three-Dimensional Neuronal Cultures-
Free pre-print version: Loading...
Abstract: Neuromodulation has made great strides in recent years, but in vitro studies have been limited to two-dimensional cell cultures, far from in vivo conditions. In this study, we realized a novel thermoplasmonic platform for modulating the neural activity of three-dimensional cell cultures, providing a new tool to bring in vitro neuromodulation studies into a three-dimensional environment. The photosensitive scaffold, obtained by covering soda-lime glass microbeads (diameter about 40 µm) with gold nanorods, integrates microbeads’ structural properties, intended to support the development of the neural network in three dimensions, with the photothermal properties of plasmonic nanoparticles. We demonstrate its efficiency in providing support for the construction of three-dimensional cell culture and how, under Near-infrared laser irradiation, their photothermal effect can precisely and non-invasively modulate the activity of the neural network. Our platform is expected to be a useful tool for improving neural network studies to better understand complex brain functions and neural disorders.
PubDate: 2022-09-13
DOI: 10.1007/s13206-022-00082-z
-
- A Lateral Flow Assay for Nucleic Acid Detection Based on Rolling Circle
Amplification Using Capture Ligand-Modified Oligonucleotides-
Free pre-print version: Loading...
Abstract: We introduce a lateral flow assay (LFA) integrated with a modified isothermal nucleic acid amplification procedure for rapid and simple genetic testing. Padlock probes specific for the target DNA were designed for ligation, followed by rolling circle amplification (RCA) using capture ligand-modified oligonucleotides as primers. After hybridization with detection linker probes, the amplified target DNA is flowed through an LFA membrane strip for binding of gold nanoparticles as the substrate for colorimetric detection. We established and validated the “RCA-LFA” method for detection of mecA, the antibiotic resistance gene for methicillin-resistant Staphylococcus aureus (MRSA). The assay was optimized using various concentrations of primers and probes for RCA and LFA, respectively. The sensitivity was determined by performing RCA-LFA using various amounts of mecA target DNA, showing a detection limit of ~ 1.3 fmol. The specificity of the assay was examined using target DNAs for other resistance genes as the controls, which demonstrated positive detection signals only for mecA DNA, when added either individually or in combinations with the control targets. Furthermore, applying the RCA-LFA method using specifically designed probes for RNA-dependent RNA polymerase (RdRp) and receptor binding domain (RBD) gene for SARS-CoV-2, which demonstrated feasibility of the method for viral gene targets. The current method suggests a useful platform which can be universally applied for various nucleic acid targets, allowing rapid and sensitive diagnosis at point-of-care.
PubDate: 2022-09-06
DOI: 10.1007/s13206-022-00080-1
-
