Amplification System Research Articles

Kinetics-accelerated one-step detection of MicroRNA through spatially localized reactions based on DNA tile self-assembly

The localization of isothermal amplification systems has elicited extensive attention due to the enhanced reaction kinetics when detecting ultra-trace small-molecule nucleic acids. Therefore, the seek for an appropriate localization cargo of spatially confined reactions is urgent. Herein, we have developed a novel approach to localize the catalytic hairpin assembly (CHA) system into the DNA tile self-assembly nanostructure. Thanks to the precise programming and robust probe loading capacity, this strategy achieved a 2.3 × 105-fold higher local reaction concentration than a classical CHA system with enhanced reaction kinetics in theory. From the experimental results, this strategy could reach the reaction plateau faster and get access to a magnified effect of 1.57–6.99 times higher in the linear range of microRNA (miRNA) than the simple CHA system. Meanwhile, this strategy satisfied the demand for the one-step detection of miRNA in cell lysates at room temperature with good sensitivity and specificity. These features indicated its excellent potential for ultra-trace molecule detection in clinical diagnosis and provided new insights into the field of bioassays based on DNA tile self-assembly nanotechnology.

Design and analysis of high-speed data optical fiber communication system through EDFA amplifier system

Abstract This study presented design and analysis of high-speed data optical fiber communication system through EDFA amplifier system. The design with practical data for parameters to maximize the transmission bit-rate is clarified. The bit rate distance product can be optimized for the improvement of performance of high-speed long haul repeater spacing with amplification technique. The basic amplification system for optical fiber communication system is clarified. Transmitter/receiver components with the circuit connections and the operation are also indicated in details. The practical circuit model of the transmitter and receiver are designed clearly in this work. The design of distribution maps to measure the Q factor and minimum bit error rate and many different simulation models for applied to achieve high-speed data optical fiber communication system.

Optimization design of two-stage amplification micro-drive system without additional motion based on particle swarm optimization algorithm

With the increasing requirements of precision mechanical systems in electronic packaging, ultra-precision machining, biomedicine and other high-tech fields, it is necessary to study a precision two-stage amplification micro-drive system that can safely provide high precision and a large amplification ratio. In view of the disadvantages of the current two-stage amplification and micro-drive system, such as poor security, low motion accuracy and limited amplification ratio, an optimization design of a precise symmetrical two-stage amplification micro-drive system was completed in this study, and its related performance was studied. Based on the guiding principle of the flexure hinge, a two-stage amplification micro-drive mechanism with no parasitic motion or non-motion direction force was designed. In addition, the structure optimization design of the mechanism was completed using the particle swarm optimization algorithm, which increased the amplification ratio of the mechanism from 5 to 18 times. A precise symmetrical two-stage amplification system was designed using a piezoelectric ceramic actuator and two-stage amplification micro-drive mechanism as the micro-driver and actuator, respectively. The driving, strength, and motion performances of the system were subsequently studied. The results showed that the driving linearity of the system was high, the strength satisfied the design requirements, the motion amplification ratio was high and the motion accuracy was high (relative error was 5.31%). The research in this study can promote the optimization of micro-drive systems.

Clear Disclosure of Hierarchical Chirality Transfer Mechanism and Wide Full‐Color and White‐Light CPL Emissions with Both High glum and Intensity by TPE Helicates

AbstractChirality transfer and amplification play key role on research of chiral materials, but it is still a challenge to clearly disclose the chirality transfer mechanism. Here, tetraphenylethylene helicate (TPEH) with aggregation‐induced emission enhancement effect is doped into achiral nematic liquid crystal (NLC) to furnish a system of excellent chirality transfer and amplification for the first time. It is found that left‐handed helical (M) TPEH molecule with four phenyl rings sloping right‐down makes a staggered overlap with other M‐TPEH at right‐up direction for attenuating the repulsive force between phenyl rings, which is further overlapped into a right‐handed helix (P) and then drives NLC array layers to the P‐LC phase. Consequently, M‐TPEH molecule leads to P‐LC phase, while P‐TPEH arouses M‐LC phase. Remarkably, this chirality transfer can amplify the circularly polarized luminescence (CPL) of TPEH by 100‐fold and give a large absolute CPL dissymmetric factor (glum) of 0.21. Moreover, the chirality can be even transferred to other achiral fluorescence dyes, and both high absolute glum of up to 0.37 and high fluorescence quantum yield (Φf) of 92% are obtained. Consequently, a customized wide full‐color and white‐light CPL emission from violet to near infrared which is closer to natural light is obtained.

Magnetic bead-based separation of sperm cells from semen-vaginal fluid mixed stains using an anti-ACRBP antibody.

Forensic DNA analysis of semen-vaginal fluid mixed stains is essential and necessary in sexual assault cases. Here, we used a magnetic bead conjugated acrosin binding protein (ACRBP) antibody to separate and enrich sperm cells from mixed stains. Previously, western blotting indicated that ACRBP was specifically expressed in sperm cells, but not in female blood and epithelial cells, while immunofluorescence data showed ACRBP was localized to the acrosome in sperm cells. In our study, sperm were separated from mixed samples at three sperm cell/female buccal epithelial cell ratios (103:103; 103:104; and 103:105) using a magnetic bead conjugated ACRBP antibody. Subsequently, 23 autosomal short tandem repeat (STR) loci were amplified using the Huaxia™ Platinum PCR Amplification System and genotyped using capillary electrophoresis. The genotyping success rate for STR loci was 90% when the sperm to female buccal epithelial cell ratio was > 1:100 in mixed samples. Our results suggest that the magnetic bead conjugated ACRBP antibody is effective for isolating sperm cells in sexual assault cases.

Few-cycle 65-µJ pulses at 11.4 µm for ultrafast nonlinear longwave-infrared spectroscopy

Low-energy excitations can provide insight into the basic ultrafast nonequilibrium dynamics of condensed matter. High-energy femtosecond pulses in the long-wavelength infrared are required to induce such processes, and can be generated in an optical parametric chirped pulse amplification (OPCPA) system comprising three GaSe stages. A femtosecond Cr:ZnS laser serves as the front-end, providing the seed for the 2.0-µm pump and the 2.4-µm signal pulses without nonlinear conversion processes. The OPCPA system is pumped at 2.05 µm by a picosecond Ho:YLF regenerative amplifier at a 1-kHz repetition rate. The recompressed idler pulses at 11.4 µm have a duration of 185 fs and an unprecedented energy of 65 µJ, corresponding to a pump-to-idler conversion efficiency of 1.2%. Nonlinear transmission experiments in the range of the L2 infrared band of liquid water demonstrate the potential of the pulses for nonlinear vibrational spectroscopy of liquids and solids.

Allelic and haplotypic polymorphisms and paternal genetic analysis of Chinese Shaanxi Han population utilizing a multiplex Y-STR set

Background The analysis of Y chromosomal genetic markers is of great significance in human genetic fields related to male individuals. The Han nationality is the most populous ethnic group. It is critical to investigate the Y-chromosome short tandem repeat (Y-STR) genetic informativeness of Han nationalities in different Chinese regions in order to gain a comprehensive understanding of their paternal genetic relationships and origin. Aim To assess the allelic and haplotypic polymorphisms of the novel AGCU Y SUPP STR amplification system containing seven Y-STRs in the maximal dataset of the Y-STR Haplotype Reference Database (YHRD) and 17 newly included Y-STRs, and explore the genetic relationships among the Shaanxi Han population and 12 reference populations from China. Subjects and methods A total sample of 220 Han male subjects were obtained from the Shaanxi Province, China, and genotyped by the novel AGCU Y SUPP STR amplification system. Multiplex population genetic analyses derived from the same 16 Y-STR loci were carried out among the Shaanxi Han population and 12 reference populations from China. Results The gene diversities (GD) ranged from the maximum value of 0.9609 (DYS385a,b) to the minimum value of 0.5441 (DYS531). Besides, 217 distinct haplotypes were detected wholly in 220 individuals, of which 214 (98.62%) were exclusive. The entire haplotype diversity (HD) and discrimination capacity (DC) were 0.9999 and 0.9864, respectively, while the haplotype match probability (HMP) was 0.0045. Among the reference populations, the obtained results of population genetic analyses revealed that the Shaanxi Han population had the largest genetic distance with the Guangxi Yao group, but the smallest genetic distance with the Hunan Tujia group. Conclusions These Y-STR loci in the AGCU Y SUPP STR amplification system were of high genetic polymorphisms and the amplification system could be used as a prospective complementary tool for forensic application and paternal genetics in the Shaanxi Han population.

Development of large-scale electret fabrication system for triboelectric nanogenerator electrical output amplification

The triboelectric nanogenerator (TENG), which converts mechanical energy to electrical output, is garnering attention owing to their application in low-energy electronic devices as a power source. To utilize the TENG as a power source, amplifying the electrical output and enhancing energy conversion efficiency are necessary. As the TENG generates electrical output through contact electrification and electrostatic charge induction, the electrical output can be amplified by enhancing the two mechanisms through the enlargement of the TENG area and the adoption of the pre-charge injected film (electret). In this regard, the fabrication of the large-scale electret is significant for amplifying the electrical output of the TENG. In this study, a corona charging system is proposed for the fabrication of the large-scale electret with a uniform charge-distributed area. Corona charging injects O2 ions into a dielectric film with an electric field generated between the pin electrode and ground electrode (GE). Through the utilization of a multi-PE (MPE) and conveyor belt-shaped GE (CBsGE) in the corona charging system, the O2 molecules near the pins can be ionized simultaneously while the CBsGE is rotating. Thus, the O2 ions can be injected uniformly into a wide area. About a voltage of 400 V and current of 45 μA can be generated by applying the fabricated electret to the TENG. As several electrets with various sizes can be fabricated simultaneously, the TENG with high electrical output can be batch fabricated to enhance its practicality significantly.

Efficient generation of femtosecond millijoule pulses at 3.1 µm.

3.2-mJ, 92-fs pulses centered at 3.1 µm are generated at a 1-kHz repetition rate through a tabletop optical parametric chirped pulse amplification (OPCPA) system based on ZnGeP2 crystals. Pumped by a 2-µm chirped pulse amplifier with a flat-top beam profile, the amplifier achieves a 16.5% overall efficiency, which, to the best of our knowledge, is the highest efficiency achieved by OPCPA at this wavelength. Harmonics up to the seventh order are observed after focusing the output in the air.

Validation of the Assurance® GDS for Cronobacter Tq II in Infant Formulas, Infant Cereals, Ingredients, and Environmental Samples Collaborative Study: First Action 2021.08.

The Assurance® GDS for Cronobacter Tq II assay is a nucleic acid amplification system for the qualitative detection of Cronobacter. The method uses an upfront concentration of the target organism from the enrichment by immunomagnetic separation (IMS) using the PickPen®device. The Assurance GDS for Cronobacter Tq II method was evaluated for Official Methods of AnalysisSM certification. The matrix was compared to the ISO 22964:2017: Microbiology of the Food Chain-Horizontal Method for the Detection of Cronobacter spp. standard and using an alternative confirmation procedure. The alternative method was evaluated using 10 g test portions in an unpaired study design for powdered infant formula (milk based with iron and DHA) containing probiotics. Eleven technicians from eight laboratories located within the United States and Europe participated in the collaborative study. Statistical analysis was conducted according to the probability of detection (POD) statistical model as presented in the AOAC validation guidelines. The difference in laboratory POD (dLPODC) values with 95% confidence intervals across collaborators was calculated for each level between the candidate and reference method results and between the candidate presumptive and confirmed results. Results obtained for the low inoculum level test portions produced a dLPOD value with a 95% confidence interval of 0.03 (-0.18, 0.15). The dLPOD results indicate equivalence between the candidate method and reference method for the matrix evaluated. The method also demonstrated acceptable inter-laboratory reproducibility as determined in the collaborative evaluation. There were no false negative results; the false positive rate was determined and produced a value of <2%. Based on the data generated, the method demonstrated Assurance GDS for Cronobacter Tq II assay produced acceptable interlaboratory reproducibility data and statistical analysis. The Assurance GDS for Cronobacter Tq II method is suitable for the rapid qualitative detection of Cronobacter in infant formulas, infant cereals, ingredients, and environmental samples.

Intermolecular and Intramolecular Priming Co-directed Synergistic Multi-strand Displacement Amplification Empowers Ultrasensitive Determination of microRNAs.

The construction of isothermal nucleic acid amplification systems with extremely high signal amplification abilities is of great importance for biomarker detection and disease diagnosis. Herein, for the first time, we proposed an intermolecular and intramolecular priming co-directed synergistic multi-strand displacement amplification (SM-SDA) system for microRNA (miRNA) detection. Strategically, the SM-SDA system is made of a single multi-functionalized hairpin probe (MF-HP) that is engineered with a long stem and endowed with target complementation, configuration transformation, enzyme recognition, and signal reporting abilities. The presence of a specific target, the model of miRNA-21, to react with MF-HP would cause an intermolecular priming-directed strand replication in parallel with an intramolecular priming-directed strand replication. The co-directed priming pattern results in the occurrence of SM-SDA containing a target-induced cyclical strand displacement amplification (T-CSDA), a target analogue primer (TAP)-induced CSDA (TAP-CSDA), and a nicked trigger primer (NTP)-induced CSDA (NTP-CSDA). The resulting multiple circuits with a synergetic signal amplification capacity remarkably enhance the target miRNA response, which has not only improved the sensitivity for qualitative and quantitative detection of miRNA but also realized the analysis of target miRNA from real clinical samples. As a proof-of-concept study, this simple but attractive isothermal signal amplification system holds a great potential for molecular diagnosis of diseases and will stimulate interest, new ideas, and discoveries in this fascinating field.

Development of a multiplex panel with 31 multi-allelic InDels for forensic DNA typing.

Insertion/Deletion (InDel) polymorphic genetic markers are abundant in human genomes. Diallelic InDel markers have been widely studied for forensic purposes, yet the low polymorphic information content limits their application and current InDel panels remain to be improved. In this study, multi-allelic InDels located out of low complexity sequence regions were selected in the datasets from East Asian populations, and a multiplex amplification system containing 31 multi-allelic InDel markers and the Amelogenin marker (FA-HID32plex) was constructed and optimized. The preliminary study on sensitivity, species specificity, inhibitor tolerance, mixture resolution, and the detection of degraded samples demonstrates that the FA-HID32plex is highly sensitive, specific, and robust for traces and degraded samples. The combined power of discrimination (CPD) of 31 multi-allelic InDel markers was 0.999 999 999 999 999 999 85, and the cumulative probability of exclusion (CPE) was 0.999 920 in a Chinese Han population, which indicates a high discrimination power. Altogether, the FA-HID32plex panel could provide reliable supplements or stand-alone information in individual identification and paternity testing, especially for challenging samples.

High sensitivity SARS-CoV-2 detection using graphene oxide-multiplex qPCR

The emerging pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) critically challenges early and accurate virus diagnoses. However, the current gold standard for SARS-CoV-2 detection, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), has reportedly failed to detect low-viral loads. One compound, graphene oxide (GO), which adsorbs single-stranded DNA (ssDNA), has been widely applied in molecular pathogen detection. This study presents a highly sensitive GO-multiplex qPCR method for simultaneous detection of two SARS-CoV-2 genes (RdRP and E) and one reference gene (RNase P). In a GO-multiplex qPCR system, GO pre-absorbs each forward primer to form specific GO-forward primer composites before entering the amplification system. Target gene amplification is confined within the primer-enriched composites, thus, improving the sensitivity of the assay. Compared to conventional multiplex qPCR, GO-multiplex qPCR reduces the limit of detection by 10-fold to 10 copies/reaction. Hence, the GO-multiplex qPCR assay can be effectively used for SARS-CoV-2 detection.

Experimental study on the fluctuation–magnification effect of chirped volume Bragg grating compressor in high-energy, ultrafast, thin-disk chirped pulse amplification system

The fluctuation–magnification effect on the peak intensity of a laser pulse caused by the nonlinear Kerr effect in the chirped volume Bragg grating (CVBG) compressor is investigated experimentally for a high-energy, thin-disk, chirped pulse amplification system. The nonlinear Kerr effect occurs at the blue end, and the accumulated nonlinear phase shift (B-integral) in the CVBG rises with the increase of laser pulse energy. Experiments show that small fluctuations in peak power of uncompressed pulses cause increasing of the temporal fluctuation and spatial fluctuation due to high Kerr-nonlinearity in the CVBG when B-integral is larger than π. Thus the initial fluctuation would be magnified by the CVBG compressor.

Kinetics of Drug Molecule Interactions with a Newly Developed Nano-Gold-Modified Spike Protein Electrochemical Receptor Sensor.

In March 2020, the World Health Organization (WHO) declared COVID-19 a pandemic, and the spike protein has been reported to be an important drug target for anti-COVID-19 treatment. As such, in this study, we successfully developed a novel electrochemical receptor biosensor by immobilizing the SARS-CoV-2 spike protein and using AuNPs-HRP as an electrochemical signal amplification system. Moreover, the time-current method was used to quantify seven antiviral drug compounds, such as arbidol and chloroquine diphosphate. The results show that the spike protein and the drugs are linearly correlated within a certain concentration range and that the detection sensitivity of the sensor is extremely high. In the low concentration range of linear response, the kinetics of receptor–ligand interactions are similar to that of an enzymatic reaction. Among the investigated drug molecules, bromhexine exhibits the smallest Ka value, and thus, is most sensitively detected by the sensor. Hydroxychloroquine exhibits the largest Ka value. Molecular docking simulations of the spike protein with six small-molecule drugs show that residues of this protein, such as Asp, Trp, Asn, and Gln, form hydrogen bonds with the -OH or -NH2 groups on the branched chains of small-molecule drugs. The electrochemical receptor biosensor can directly quantify the interaction between the spike protein and drugs such as abidor and hydroxychloroquine and perform kinetic studies with a limit of detection 3.3 × 10−20 mol/L, which provides a new research method and idea for receptor–ligand interactions and pharmacodynamic evaluation.

A Highly Sensitive Fluorescence and Screen-Printed Electrodes-Electrochemiluminescence Immunosensor for Ricin Detection Based on CdSe/ZnS QDs with Dual Signal.

A sensitive dual-readout immunosensor for fluorescence and electrochemiluminescence (ECL) detection of ricin was established, which was combined with a streptavidin–biotin signal amplification system. CdSe/ZnS quantum dots with fine fluorescence and ECL properties were used as the dual-signal function probes of the sandwich immunocomplex. Under the optimum experimental conditions, the dual signal intensity increased significantly with the rise in ricin concentration. The fluorescence intensity of the senor exhibited a good liner relationship toward the ricin concentrations with 0.1~100 ng/mL and the limit of detection (LOD) was 81.7 pg/mL; taking ECL as the detection signal, the sensor showed a linear relationship with the ricin concentrations ranging from 0.01 ng/mL to 100 ng/mL and the LOD was 5.5 pg/mL. The constructed sensor with high sensitivity had been successfully applied to the detection of ricin in complex matrices with satisfactory recoveries. The proposed immunosensor model can be extended to the analysis and detection of others target proteins.

The study of the performance of a microbial fuel cell: a progress towards the improvement of low electrical bioenergy output by using an amplification system

A microbial fuel cell (MFC) has been conceived and constructed for the treatment of the sheep manure wastes and their conversion into clean sustainable renewable energy. The aim of the present investigation was to examine the performance of this bioelectrochemical device, in breaking down the organic matter (pollutant removal) and simultaneously producing electricity. Furthermore, the objective was to enhance the low electric energy by using an adequate amplification system. So, the chemical oxygen demand (COD) removal was increased by 58.7% with the MFC running for 10days. However, this technology faces practical barriers as it produces low electrical energy. A power management system was therefore elaborated in this respect. It included the MFC, operational amplifier (OA), solar photovoltaic panel and a boost DC/DC converter. The low voltage output obtained was thus increased substantially using the OA prior to its polarization by the solar photovoltaic module. The amplified voltage was sufficiently enough and in consequence, utilized to feed a light emitting diode. The low output voltage 0.5V was simply harvested, successfully boosted up to approximately 2V (i.e. 4 times higher) and finally harnessed as a power supply. The MFCs association shows the positive stacking effect successfully, when the cells were connected in parallel. This novel application is very interesting to utilize the natural bioenergy contained in wastes to supply small electronic devices.

An Ultrasensitive Electrochemical Biosensor Integrated by Nicking Endonuclease-Assisted Primer Exchange Reaction Cascade Amplification and DNA Nanosphere-Mediated Electrochemical Signal-Enhanced System for MicroRNA Detection.

Specific and sensitive microRNAs (miRNAs) detection is essential to early cancer diagnosis. The development of these technologies including functional nuclease-mediated target amplification and DNA nanotechnology possesses tremendous potential for the high-performance detection of miRNAs in the accurate diagnosis of disease. In this study, we have established an ultrasensitive electrochemical biosensor by combining nicking endonuclease-assisted primer exchange reaction (PER) cascade amplification with a DNA nanosphere (DNS)-mediated electrochemical signal-enhanced system for the detection of miRNA-21 (miR-21). The cascade amplification is initiated by a nicking endonuclease that can cleave specific DNA substrates and highly amplify translation of the target to single-stranded DNA fragments (sDNA). Then, the PER cascade is powered by strand-displacing polymerase and generates a large amount of nascent single-stranded connector DNA (cDNA) via sDNA triggering of the dumbbell probe (DP), thus achieving the cascade amplification of the target. Finally, the DNS loaded with plenty of electroactive substances can be captured on the electrode via cDNA for further enhancing the electrochemical signal and highly sensitive detection of miR-21. The proposed electrochemical biosensor exhibits a wide detection range of 1 aM to 0.1 nM and a low detection limit of 0.58 aM. The excellent selectivity allows the biosensor to discriminate miR-21 from other miRNAs, even the one base-mismatched sequence. Moreover, the practicability of the biosensor is investigated by analyzing miR-21 in human serum and cancer cell lysate. Therefore, our proposed nicking endonuclease-assisted PER cascade amplification strategy provides a powerful platform for the early detection of miRNA-related disease and molecular diagnosis.

Design and Optimization of Amplifying Mechanism in Gas-Liquid Thermoelectric Power Device

Aiming at the small output displacement of the gas-liquid temperature difference power device, a flexible hinge displacement amplification system was designed and optimized using the lever principle. The formula for calculating the magnification of the lever-type flexible hinge mechanism was derived by taking into account the offset of the flexible hinge's rotational center. Likewise, consider how the parameters of the mechanism affect the output performance of the amplifying mechanism. Combining SolidWorks and Workbench to improve the amplifying mechanism's parameters, find the best value for the parameters, and experimentally validate the optimized results. The research shows that the error between the experimental data and the finite element simulation value is 2.49%, which verifies the correctness of the simulation model. The design goals of high magnification of the mechanism and output after displacement amplification are realized.

Triple quantitative detection of three inflammatory biomarkers with a biotin-streptavidin-phycoerythrin based lateral flow immunoassay

Quantified inflammatory biomarkers are effective clinical strategy for correct and reasonable drug treatment. In the study, a triple lateral flow immunoassay (triple LFIA) had firstly been developed for specific and simultaneous detection of three pivotal inflammatory biomarkers (PCT, CRP and SAA) via biotin-streptavidin-phycoerythrin signal amplification system in one strip. The developed triple LFIA adopted phycoerythrin (PE) as chromophore to eliminate auto-fluorescence interference from plasma biomolecules and anti-PE mAb as single control line to reduce the nonspecific adsorption, which featured particular advantages in high sensitivity and specificity in a large range of analyte concentrations with the LODs of 0.106 ng/mL for PCT, 0.345 μg/mL for CRP and 3.112 μg/mL SAA, respectively. And the linear quantitative detection ranges were from 0.106 to 100 ng/mL, from 0.345 to 200 μg/mL, and from 3.112 to 200 μg/mL, respectively. Compared to commercial chemiluminescence immunoassay method, the correlations for tested PCT, CRP and SAA in 108 clinical samples were 0.989, 0.987 and 0.988, respectively. In summary, we had proposed a rapid and accurate plasma detection to measure inflammation factors, which facilitated the clinical value to achieve precise treatment.