查看更多>>摘要:The cancer biomarkers including AFP, CEA, CA199 and CA125, are of great importance in the diagnosis, prognostic prediction and recurrence monitoring of malignancies. However, in clinical practical applications, most tumor cancer biomarkers are lack of sensitivity and specificity. In this study, we propose a terahertz (THz) metasurface (MS) immunosensor coupled with gold nanoparticles (AuNPs), which have good biocompatibility and high specific surface area for biomarkers. Firstly, we added AuNPs to the surface of the sensor. And then, the surface is modified with Anti-CA125 or Anti-CA199 to improve the sensitivity and specificity to the target antigen. The biosensor was fabricated using a surface micromachining process and characterized by a THz-time-domain spectroscopy (TDS) system. The sensitivity of the resonance frequency of the sensor to the refractive index was 65 GHz/RIU (refractive index unit). The detection performance of the THz immunosensor was also verified with different concentrations of CA125 and CA199. The experimental results showed that the frequency shift of the resonance peak was linearly related to the concentration of CA125 and CA199. The detection limits for both CA125 and CA199 are 0.01 U/ml, which is better than that of other common methods. Finally, serum samples were collected and detected to explore whether this method is suitable for clinical detection. The results are consistent with the results of antigen recognition. This study proves that the practicability of the THz immunosensor, which potentially provides important techniques and equipment for improving the sensitivity and specificity of cancer biomarkers.
查看更多>>摘要:In this study, a modified solution cathode glow discharge atomic emission spectrometry (SCGD-AES) was used to detect metal elements in electroplating sewage. The SCGD-AES device was equipped with a portable semiconductor anode refrigeration unit, which was built independently. The red-heat effect of tungsten electrode was alleviated by adding the portable refrigeration unit, thus improving thermal stability with the withstand voltage from 1040 V to 1140 V. Compared with the devices without semiconductor refrigeration, the chromium was excited more favorable when the discharge voltage increased, and the limit of detection (LOD) decreased by 8.5 times. Furthermore, the LODs of Zn, Cd, Ni, Cu and Pb decreased by 1.8-3.2 times, respectively, which realized the detection of elements in electroplating sewage and showed high performance in the field of trace elements analysis. Furthermore, the accuracy of the method was verified by stream sediment reference material (GBW07312), and the results were consistent with the certified values. The recoveries of elements added to industrial sewage and seawater range were from 90.5 to 98.7%, demonstrating good accuracy of the proposed method.
查看更多>>摘要:In the extraction process of traditional Chinese medicine (TCM), the rapid detection of the active components of TCM plays a vital role. We pioneered the choice of dual-spectrum technology based on near-infrared (NIR) and ultraviolet-visible (UV-Vis) spectra and applied it to the quantitative monitoring of online extraction of puerarin and total flavonoids from Pueraria lobata. An online monitoring system was constructed to fulfill the dual spectrum online monitoring. The extraction solution flows from a flask into a pipeline under the control of a peristaltic pump, removing the solid residues contained in the solution with filters. When the extraction solution flows into the cuvette, it will be measured with NIR and UV-Vis portable spectrometers. After finishing the spectra collection, the peristaltic pump will reverse to push the extraction solution back to the flask. The partial least squares (PLS) models were established to quantitatively analyze puerarin and total flavonoids by NIR, UV-Vis, and dual-spectrum data. As for NIR, UV-Vis and dual-spectrum, the RMSEP (mg/mL) of the puerarin models were 0.0803, 0.0500, and 0.0351, and those of the total flavonoids models were 0.4373, 0.2402, and 0.1795, respectively. The results show that the dual-spectrum online monitoring system is simple, stable and fast (a few seconds) in operation, and accurate in prediction. Compared with individual NIR or UV-Vis spectra, the dual-spectra contains more comprehensive information, and the prediction ability is stronger. This method is expected to become a promising tool for the quantitative analysis of other traditional Chinese medicines.
查看更多>>摘要:A combination of Hybridization Chain Reaction (HCR) and Lateral Flow Immunoassay (LFIA) is an attractive strategy for a simple signal amplification DNA/RNA detection. The present study aimed to report a strategy used to solve a problem encountered when the target DNA contained folded secondary structure during HCR, enabling HCR hairpin probes to easily access the target site. The 24-nt conserved sequence within 3 & PRIME;-UTR, present only in dengue virus genome but not in other species, is an ideal target to use as a probe binding site for pan-dengue virus detection. Thus, the 105-nt target containing the 24-nt target sequence was chosen as a target with sec-ondary structures. The 24-nucleotide (nt) synthetic target DNA successfully induced HCR reaction within 5 min at room tem-perature. However, the HCR detection of the 105-nt synthetic target DNA with secondary structures was prob-lematic. The probe hybridization was prevented by the secondary structures of the target, resulting in a failure to generate HCR product. To solve this problem, two helper oligonucleotides (helper1 and helper2) were designed to linearize the folded structure of the 105-nt target through strand-displacement mechanism, allowing the HCR hairpin probes to easily access the target site. The HCR product with the labeled helper oligonucleotides and the labeled probes were successfully detected by LFIA. With this strategy, the combination of the helper-enhanced HCR and LFIA exhibited a limit of detection (LOD) in a nanomolar range of the 105-nt DENV synthetic target DNA. Our study demonstrated that signal amplification by the combination of HCR and LFIA could successfully detect the target DNA with secondary structure, but not target RNA with secondary structure. In summary, this work provided a proof of concept of two main issues including probe hybridization enhancement by helper oligonucleotide for the target with complicated secondary structure and the advantage of a combination of labeled helper and HCR probes design for LFIA to overcome the false positive result from HCR probe leakage. Our findings on the use of helper oligonucleotides may be beneficial for the development of other isothermal amplification, since the secondary structure of the target is one of the major obstacles among hybridization-based methods.
查看更多>>摘要:Bisphenols, parabens, and their metabolites are a group of chemical compounds with a wide range of polarities but similar chemical structures, which presents a challenge for the simultaneous determination of these compounds in complex biological samples. In this study, a rapid and sensitive method for simultaneous quantification of free bisphenol A (BPA), conjugated BPA, bisphenols, and parabens analogs was developed using solid-phase extraction (SPE) tandem liquid-liquid extraction (LLE). We compared the effects of different types of SPE cartridges, diluents, and LLE solvents on the analyte recovery. Utilizing the direct and indirect determination methods (enzyme hydrolysis), we confirmed the accuracy of the direct method for measuring BPA glucuronide and BPA disulfate. The method enabled the analysis of 24 endocrine-disrupting chemicals (EDCs) in one injection through UHPLC-MSMS measurements, with satisfactory recovery (mean: 91.8-98.6% for urine, 80.2%-96.8% for serum) and precision (RSD <15%). The LOD and LOQ values were 0.003 and 0.01 ng/mL for serum, and 0.002 and 0.006 ng/mL for urine samples, respectively. For real sample analysis, the median concentration of analytes in serum and urine samples ranged from 0.04 ng/mL (BPS) to 56.4 ng/mL (4-HB) and 0.11 ng/mL (BPA) to 136 ng/mL (4-HB), respectively. This method provides a new strategy to simultaneously identify compounds with a wide range of polarities from complicated biological matrices.
Iles, Alice H.He, Peijun J. W.Katis, Ioannis N.Horak, Peter...
8页
查看更多>>摘要:Lateral flow devices (LFDs) or lateral flow tests (LFTs) are one of the most widely used biosensor platforms for point-of-care (POC) diagnostics. The basic LFD design has remained largely unchanged since its first appearance, and this has limited LFD use in clinical applications due to a general lack of analytical sensitivity. We report here a comprehensive study of the use of laser-patterned geometric control barriers that influence the flow dynamics within an LFD, with the specific aim of enhancing LFD sensitivity and lowering the limit of detection (LOD). This control of sample flow produces an increase in the time available for optimizing the binding kinetics of the implemented assay. The geometric modification to the flow path is in the form of a constriction that is produced by depositing a photo-sensitive polymer onto the nitrocellulose membrane which when polymerized, creates impermeable barrier walls through the depth of the membrane. Both the position of the constriction within the flow path and the number of constrictions allow for an increase in the sensitivity because of a slower overall flow rate within the test and a larger volume of sample per unit width of the test line. For these high sensitivity LFDs (HS-LFD), through optimization of the constriction position and addition of a second constriction we attained a 62% increase in test line color intensity for the detection of procalcitonin (PCT) and were also able to lower the LOD from 10 ng/mL to 1 ng/mL. In addition, of relevance for future commercial exploitation, this also significantly decreases the antibody consumption per device leading to reduced costs for test production. We have further tested our HS-LFD with contrived human samples, validating its application for future clinical use.
查看更多>>摘要:In this work, strong electrochemiluminescence (ECL) emission was achieved by using one type of the G-quadruplex selective iridium (III) complex as an efficient ECL signal probe. Based on the typical sandwich immunoreaction between the cardiac troponin-I antigen (cTnI) and its corresponding antibody, iridium (III) complex was introduced according to its specific interaction with G-quadruplex DNA that modified on the surface of negatively charged gold nanoparticles ((- )AuNPs), inducing an increased ECL signal, which was proportional to cTnI concentration. Based on of this, quantitative detection of cTnI could be realized in the range of 5.0 fg/mL100 ng/mL, with a detection limit of 1.67 fg/mL. Moreover, the proposed immunosensor was successfully applied for the diagnosis of cTnI in human serums from healthy individuals and acute myocardial infarction (AMI) patients, suggesting a great potential application value in the early diagnosis of AMI.
查看更多>>摘要:The wide clinical application of D-penicillamine (D-PA) makes it inevitably accumulates in the environment, seriously threatening human health and the ecological environment. To better supervisory control D-PA, a highly sensitive and reliable photoelectrochemical (PEC) sensor based on gold nanoparticles (Au NPs) loaded on graphitic carbon nitride sheet and hexagonal NH2-UiO-66 composite (g-C3N4/Au/NH2-UiO-66 ) was synthesized. Tactfully using the strong bonding between D-PA and Au NPs and the effective carrier separation of Z-scheme heterojunction, the designed g-C3N4/Au/NH2-UiO-66 PEC sensor without an extra recognition unit exhibited a selective and sensitive photocurrent to D-PA. With the aid of UV diffuse reflectance spectra (UV-DRs), electron paramagnetic resonance (EPR) characterization, and free radical capture experiments, the electron transfer path of the PEC sensing system was deduced. The proposed g-C3N4/Au/NH2-UiO-66 PEC-based sensor achieved a low detection limit of 0.0046 mu M (S/N = 3) with a wide linear response ranging from 10 nM to 400 mu M. In addition, its good stability and selectivity also laid a good foundation for practical applications.
查看更多>>摘要:Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a serious threat to human health. Current methods such as reverse transcription polymerase chain reaction (qRT-PCR) are complex, expensive, and time-consuming. Rapid, and simple screening methods for the detection of SARS-CoV-2 are critically required to fight the current pandemic. In this work we present a proof of concept for, a simple optical sensing method for the screening of SARS-CoV-2 through its spike protein subunit S1. The method utilizes a target-specific extractor chip to bind the protein from the biological specimens. The disulfide bonds of the protein are then reduced into a biothiol with sulfhydryl (SH) groups that react with a blue-colored benzothiazole azo dye-Hg complex (BAN-Hg) and causes the spontaneous change of its blue color to pink which is observable by the naked eye. A linear relationship between the intensity of the pink color and the logarithm of reduced S1 protein concentration was found within the working range 130 ng.mL(-1) -1.3 pg mL(-1). The lowest limit of detection (LOD) of the assay was 130 fg mL(-1). A paper based optical sensor was fabricated by loading the BAN-Hg sensor onto filter paper and used to screen the S1 protein in spiked saliva and patients' nasopharyngeal swabs. The results obtained by the paper sensor corroborated with those obtained by qRT-PCR. The new paper-based sensing method can be extended to the screening of many viruses (e.g. the human immunodeficiency virus, the human polyomavirus, the human papilloma virus, the adeno associated viruses, the enteroviruses) through the cysteine residues of their capsid proteins. The new method has strong potential for screening viruses at pathology labs and in remote areas that lacks advanced scientific infrastructure. Further clinical studies are warranted to validate the new sensing method.