highly sensitive

  • 文章类型: Journal Article
    非常需要具有宽范围和高灵敏度的柔性压力传感器,但构建起来具有挑战性。在这里,我们已经成功地制造了一个压力温度双传感器通过离子辅助电荷增强策略。受益于[EMIM+][TFSI-]离子对的固定作用和离子液体(IL)与HFMO(H10Fe3Mo21O51)之间的电荷转移,所形成的IL-HFMO-TPU压力传感器具有25.35kPa-1的高灵敏度和宽传感范围(~10MPa),分别。此外,传感器装置具有高耐久性和稳定性(5000次循环@1MPa)。IL-HFMO-TPU传感器还显示出良好的温度感测特性的优点。归因于这些优越的特性,所提出的传感器设备可以检测超宽传感范围(从Pa到MPa)内的压力,包括呼吸和生物物理信号监测等。所提出的离子辅助增强方法是构建高性能柔性压力-温度双传感器的通用策略。
    Flexible pressure sensors with a broad range and high sensitivity are greatly desired yet challenging to build. Herein, we have successfully fabricated a pressure-temperature dual sensor via an ionic assisted charge enhancement strategy. Benefiting from the immobilization effect for [EMIM+] [TFSI-] ion pairs and charge transfer between ionic liquid (IL) and HFMO (H10Fe3Mo21O51), the formed IL-HFMO-TPU pressure sensor shows a high sensitivity of 25.35 kPa-1 and broad sensing range (∼10 MPa), respectively. Furthermore, the sensor device exhibits high durability and stability (5000 cycles@1 MPa). The IL-HFMO-TPU sensor also shows the merit of good temperature sensing properties. Attributed to these superior properties, the proposed sensor device could detect pressure in an ultrawide sensing range (from Pa to MPa), including breathe and biophysical signal monitoring etc. The proposed ionic assisted enhancement approach is a generic strategy for constructing high performance flexible pressure-temperature dual sensor.
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  • 文章类型: Journal Article
    COVID-19大流行强调了快速、敏感,以及检测大量呼吸道病毒的有效方法。检测呼吸道病毒的“黄金标准”分子检测,如定量聚合酶链反应(qPCR)和逆转录qPCR(RT-qPCR),依赖于侵入性拭子样品,需要耗时和劳动密集型的提取过程。此外,基于RT-qPCR的分析的周转时间对于快速筛选来说太长。基于无提取唾液的方法提供了具有快速周转时间的非侵入性取样过程,并且适合于高通量应用。然而,当与标准RT-qPCR系统一起使用时,没有提取显著降低了测定的灵敏度。这里,使用一种新颖的光调制生物传感(OMB)平台,我们开发了一种快速且高度灵敏的无唾液提取分子检测方法.我们盲目地测试了364对来自以色列疑似SARS-CoV-2病例的鼻咽拭子和唾液样本。与基于金标准拭子的RT-qPCR检测相比,基于无提取唾液的OMB测定的灵敏度为90.7%,比基于无唾液提取的RT-qPCR检测的灵敏度(77.8%)高得多,具有相似的特异性(95.3%和97.6%,分别)。此外,在通过基于OMB的检测鉴定为阳性的12个样本中,从COVID-19病房的住院患者中收集了8个样本,在入院时被证实为SARS-CoV-2阳性,表明OMB检测的实际临床敏感性和特异性更高。考虑到其基于唾液的用户友好协议,短且具有成本效益的无提取过程,和高临床准确性,基于OMB的分子检测方法非常适合对大量呼吸道病毒进行高通量检测.
    目标:SARS-CoV-2爆发三年后,没有结合低成本和简单样品制备的分子测试,有效的样品处理,最少的试剂和一次性要求,高灵敏度,和大规模筛选所需的高通量。现有的快速分子技术通常牺牲某些要求来满足其他要求。然而,新型病毒性疾病的局部爆发每天都在世界不同地区发生。在这种情况下,呼吸系统疾病特别重要,因为它们经常通过空气传播并且具有高度传染性,具有快速全球传播的潜力。广泛接受的意见是,另一场大流行只是时间问题。为了确保对即将到来的“疾病X”的遏制努力取得成功,引入快速,高通量,和高度敏感的诊断方法来检测和识别病原体是至关重要的。大流行几个月后,唾液被建议作为SARS-CoV-2检测的诊断基质。唾液的收集不需要拭子并且是微创的。特别是,基于唾液的无提取化验需要更少的试剂和一次性用品,因此更快,更便宜,为低收入国家提供了一个有吸引力的替代方案。不幸的是,当前基于无唾液提取的检测方法,如直接RT-qPCR或等温扩增,具有低灵敏度或低吞吐量。因此,我们相信,提出的高度敏感的ht-OMBi平台和无提取的基于唾液的分子检测可以成为传染病监测工具箱中的重要工具。
    The COVID-19 pandemic highlighted the necessity of fast, sensitive, and efficient methods to test large populations for respiratory viruses. The \"gold standard\" molecular assays for detecting respiratory viruses, such as quantitative polymerase chain reaction (qPCR) and reverse transcription qPCR (RT-qPCR), rely on invasive swab samples and require time-consuming and labor-intensive extraction processes. Moreover, the turnaround time for RT-qPCR-based assays is too lengthy for rapid screening. Extraction-free saliva-based methods provide a non-invasive sampling process with a fast turnaround time and are suitable for high-throughput applications. However, when used with a standard RT-qPCR system, the absence of extraction significantly reduces the assays\' sensitivity. Here, using a novel optical modulation biosensing (OMB) platform, we developed a rapid and highly sensitive extraction-free saliva-based molecular assay. We blindly tested 364 paired nasopharyngeal swabs and saliva samples from suspected SARS-CoV-2 cases in Israel. Compared with the gold standard swab-based RT-qPCR assay, the sensitivity of the extraction-free saliva-based OMB assay is 90.7%, much higher than the sensitivity of extraction-free saliva-based RT-qPCR assay (77.8%) with similar specificity (95.3% and 97.6%, respectively). Moreover, out of 12 samples identified by the OMB-based assay as positive, 8 samples were collected from hospitalized patients in a COVID-19 ward and were verified to be SARS-CoV-2-positive upon admission, indicating that the actual clinical sensitivity and specificity of the OMB assay are higher. Considering its user-friendly saliva-based protocol, short and cost-effective extraction-free process, and high clinical accuracy, the OMB-based molecular assay is very suitable for high-throughput testing of large populations for respiratory viruses.
    OBJECTIVE: Three years after the SARS-CoV-2 outbreak, there are no molecular tests that combine low-cost and straightforward sample preparation, effective sample handling, minimal reagent and disposable requirements, high sensitivity, and high throughput required for mass screening. Existing rapid molecular techniques typically sacrifice certain requirements to meet others. Yet, localized outbreaks of novel viral diseases happen daily in different parts of the world. In this context, respiratory diseases are of specific importance, as they are frequently airborne and highly contagious, with the potential for a rapid global spread. The widely accepted opinion is that another pandemic is just a question of time. To ensure that the containment efforts for the upcoming \"disease X\" are successful, introducing rapid, high-throughput, and highly sensitive diagnostic methods for detecting and identifying pathogens is critical. A few months into the pandemic, saliva was suggested as a diagnostic matrix for SARS-CoV-2 detection. The collection of saliva does not require swabs and is minimally invasive. In particular, extraction-free saliva-based assays require fewer reagents and disposables, and therefore are faster and cheaper, offering an appealing alternative for low-income countries. Unfortunately, current extraction-free saliva-based detection methods, such as direct RT-qPCR or isothermal amplification, have either low sensitivity or low throughput. Therefore, we believe that the presented highly sensitive ht-OMBi platform and the extraction-free saliva-based molecular assay can become an essential tool in the infectious disease monitoring toolbox.
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  • 文章类型: Journal Article
    由于溶液中不同数量的白细胞(WBC)具有细胞的适应性渗透压,WBC本身和溶液中的浓度相似,导致它们具有相似的介电性能。因此,当WBC在溶液中时,微波传感器可能难以感测数量变化。本文提出了一种高度敏感的,用于WBC的线性介电常数启发微波生物传感器,通过蒸发法计数。提出了这种测量方法以记录细胞溶液滴落到芯片上并自然完全蒸发之后的测量结果。拟议的生物传感器由使用集成无源制造技术在GaAs衬底上制造的空气桥接非对称差分电感器和位于中心的圆形叉指电容器组成。它被优化为具有更大的敏感区域和改进的Q因子,这增加了细胞与电磁场之间相互作用的有效面积,并有助于检测它们的数量变化。传感依赖于细胞的介电特性和不同浓度的介电常数的变化,以及共振特性的变化,主要代表频率偏移,对应于细胞浓度的宏观变化。微波生物传感器用于在温度(26.00±0.40°C)和湿度(54.40±3.90RH%)环境中测量浓度为0.25×106至8×106个细胞/mL的生物样品。测量结果表明,高灵敏度为25.06Hz/cells·mL-1,高度线性响应为r2=0.99748。此外,对悬浮中的单个细胞进行数学建模,以估计单个细胞的介电常数,并进一步解释所提出的微波生物传感器的工作机理。
    Since different quantities of white blood cells (WBCs) in solution possess an adaptive osmotic pressure of cells, the WBCs themselves and in solution have similar concentrations, resulting in them having similar dielectric properties. Therefore, a microwave sensor could have difficulty in sensing the quantity variation when WBCs are in solution. This paper presents a highly sensitive, linear permittivity-inspired microwave biosensor for WBCs, counting through the evaporation method. Such a measurement method is proposed to record measurements after the cell solution is dripped onto the chip and is completely evaporated naturally. The proposed biosensor consists of an air-bridged asymmetric differential inductor and a centrally located circular fork-finger capacitor fabricated on a GaAs substrate using integrated passive fabrication technology. It is optimized to feature a larger sensitive area and improved Q-factor, which increases the effective area of interaction between cells and the electromagnetic field and facilitates the detection of their changes in number. The sensing relies on the dielectric properties of the cells and the change in the dielectric constant for different concentrations, and the change in resonance properties, which mainly represents the frequency shift, corresponds to the macroscopic change in the concentration of the cells. The microwave biosensors are used to measure biological samples with concentrations ranging from 0.25 × 106 to 8 × 106 cells per mL in a temperature (26.00 ± 0.40 °C) and humidity (54.40 ± 3.90 RH%) environment. The measurement results show a high sensitivity of 25.06 Hz/cells·mL-1 with a highly linear response of r2 = 0.99748. In addition, a mathematical modeling of individual cells in suspension is performed to estimate the dielectric constant of individual cells and further explain the working mechanism of the proposed microwave biosensor.
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  • 文章类型: Journal Article
    卤化物钙钛矿由于具有优异的光电性能,在X射线检测中显示出巨大的潜力。然而,寻找一种具有成本效益和环境可持续的方法来处理报废设备仍然具有挑战性。这里,我们为报废钙钛矿X射线探测器引入了“一键重启”环保回收策略。这种方法,利用水,允许重新捕获和重复使用钙钛矿和导体材料。该过程简单明了,环保,消除了进一步化学处理的需要,净化,额外的添加剂或催化剂,复杂的设备。我们通过从回收材料中重建可穿戴电子设备的柔性钙钛矿膜,开发了一种可持续的设备循环。大规模,由无金属钙钛矿DABCO-N2H5-I3(DABCO=N-N'-二氮杂双环[2.2.2]八铵)制成的柔性膜在50V偏压下实现了6204±268μCGyair-1cm-2的显著令人印象深刻的平均灵敏度和102.3Gynairs-1的低检测限,这对X射线成像非常有效。回收柔性设备的灵敏度不仅与用新鲜材料制成的单晶设备相匹配,而且在所有无金属钙钛矿X射线探测器中排名最高。“一键重启”应用于从老化的单晶对应物衍生的可扩展柔性设备,提供巨大的成本,时间,和节能相比,它们的单晶等价物。这些优势大大增强了未来的市场竞争力。本文受版权保护。保留所有权利。
    Halide perovskites have shown great potential in X-ray detection due to outstanding optoelectronic properties. However, finding a cost-effective and environmentally sustainable method for handling end-of-life devices has remained challenging. Here, a \"One-Click Restart\" eco-friendly recycling strategy is introduced for end-of-life perovskite X-ray detectors. This method, utilizing water, allows for the recapture and reuse of both perovskite and conductor materials. The process is straightforward and environmentally friendly, eliminating the need for further chemical treatment, purification, additional additives or catalysts, and complex equipment. A sustainable device cycle is developed by reconstructing flexible perovskite membranes for wearable electronics from recycled materials. Large-scale, flexible membranes made from metal-free perovskite DABCO-N2H5-I3 (DABCO = N-N\'-diazabicyclo[2.2.2]octonium) achieve remarkably impressive average sensitivity of 6204 ± 268 µC Gyair -1 cm-2 and a low detection limit of 102.3 nGyair s-1, which makes highly effective for X-ray imaging. The sensitivity of recycled flexible devices not only matches that of single-crystal devices made with fresh materials but also ranks as the highest among all metal-free perovskite X-ray detectors. \"One-Click Restart\" applies to scalable flexible devices derived from aged single-crystal counterparts, offering significant cost, time, and energy savings compared to their single-crystal equivalents. Such advantages significantly boost future market competitiveness.
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  • 文章类型: Journal Article
    猴痘病毒(MPXV),导致传染性疾病猴痘的病原体,导致皮肤上的病变,淋巴结病,和发烧。自2022年5月以来,它已构成全球公共卫生威胁。MPXV的高灵敏度和特异性检测对于预防疾病的传播至关重要。PyroccusfuriosusArgonaute(PfAgo)是一种人工DNA指导的限制性切割酶,具有5'-磷酸化ssDNA序列,它可以被开发来特异性地检测病原体的核酸。这里,建立了基于PfAgo的系统,用于检测靶向F3L基因的MPXV特异性DNA。通过快速PCR程序可以获得79bp的短扩增子,在45分钟内完成。设计了两个5'-磷酸化指导DNA来指导PfAgo切割扩增子,以获得MPXV特异性的18bp5'-磷酸化序列,不是其他正痘病毒(牛痘,天花,和牛痘病毒)。18bp序列引导PfAgo切割设计的MPXV特异性探针以发射荧光。在PfAgo-MPXV系统的优化条件下,可以在60分钟内完成提取的MPXVDNA的检测,检测限(LOD)为1.1拷贝/反应,并且不依赖于昂贵的仪器。PfAgo-MPXV系统在模拟咽拭子中灵敏检测MPXV中的成功应用,皮肤拭子,sera,和废水证明了系统的良好性能。PfAgo平台,这里建立了高灵敏度和特异性,有可能阻止MPXV的传播。
    Monkeypox virus (MPXV), the pathogen responsible for the infectious disease monkeypox, causes lesions on the skin, lymphadenopathy, and fever. It has posed a global public health threat since May 2022. Highly sensitive and specific detection of MPXV is crucial for preventing the spread of the disease. Pyrococcus furiosus Argonaute (PfAgo) is an artificial DNA-guided restriction cleavage enzyme programmable with 5\'-phosphorylated ssDNA sequences, which can be developed to specifically detect nucleic acids of pathogens. Here, a PfAgo-based system was established for the detection of MPXV-specific DNA targeting the F3L gene. A short amplicon of 79 bp could be obtained through a fast PCR procedure, which was completed within 45 min. Two 5\'-phosphorylation guide DNAs were designed to guide PfAgo to cleave the amplicon to obtain an 18 bp 5\'-phosphorylation sequence specific to MPXV, not to other orthopoxviruses (cowpox, variola, and vaccinia viruses). The 18 bp sequence guided PfAgo to cleave a designed probe specific to MPXV to emit fluorescence. With optimized conditions for the PfAgo-MPXV system, it could be completed in 60 min for the detection of the extracted MPXV DNA with the limit of detection (LOD) of 1.1 copies/reaction and did not depend on expensive instruments. Successful application of the PfAgo-MPXV system in sensitively detecting MPXV in simulated throat swabs, skin swabs, sera, and wastewater demonstrated the system\'s good performance. The PfAgo platform, with high sensitivity and specificity established here, has the potential to prevent the spread of MPXV.
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  • 文章类型: Journal Article
    基于适体未修饰的金纳米颗粒的视觉检测实验在生物技术中显示出巨大的潜力。这里,我们报告了一个可见的,无盐和高度敏感的四环素(TC)测定基于胶态稳定的AuNP混合物,该混合物包含聚(二烯丙基二甲基氯化铵)封端的AuNP((())AuNP)和四环素特异性适体封端的AuNP(Apt封端的AuNP)。该报告的TC测定是可见的且无盐的,其在TC检测期间不需要任何盐。用裸眼,纳摩尔四环素浓度可以在20分钟内确定。四环素的检测极限低至1.0fM的浓度,检测范围为8个数量级(5×10-14M至5×10-6M)。此外,报道的TC测定对四环素的选择性也优于其他抗生素,金属阳离子,蛋白质和氨基酸。这些发现清楚地证明了所报道的TC测定用于TC检测和监测的高潜力。
    Visual detection assay based on aptamer unmodified gold nanoparticles shows great potential in biotechnology. Here, we reported a visible, salt-free and highly sensitive tetracycline (TC) assay based on a colloidally stable mixture of AuNPs that contains poly (diallyl dimethylammonium chloride) capped AuNPs ((+)AuNPs) and tetracycline-specific aptamer capped AuNPs (Apt-capped AuNPs). This reported TC assay was visible and salt-free that did not need any salt during TC detection. With naked eyes, nanomolar tetracycline concentrations could be identified within 20 min. A detection limit of tetracycline down to a concentration of 1.0 fM with a broad detection range of 8 order of magnitudes (5 × 10-14 M to 5 × 10-6 M) was reached. Furthermore, the reported TC assay also exhibited good selectivity for tetracycline over other antibiotics, metal cations, proteins and amino acids. These findings clearly demonstrated the high potential of the reported TC assay for TC detection and monitoring.
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  • 文章类型: Journal Article
    轻质且高度可压缩的材料在柔性压力感测装置中受到相当大的关注。在这项研究中,一系列多孔木材(PWs)是通过将处理时间调整为0至15h并通过H2O2进行额外氧化来从天然木材中化学去除木质素和半纤维素而产生的。所制备的表观密度从95.9至46.16mg/cm3不等的PWs倾向于形成具有改善的可压缩性的波形交织结构(在100kPa下高达91.89%应变)。由PW组装的传感器,处理时间为12h(PW-12),具有最佳的压阻-压电耦合传感特性。对于压阻特性,它具有15.14kPa-1的高应力敏感性,覆盖0.06-100kPa的宽线性工作压力范围。因为它的压电势,PW-12显示灵敏度为0.443V·kPa-1,超低频检测低至0.0028Hz,在0.41Hz下超过60,000次循环和良好的可循环性。自然衍生的全木压力传感器在满足电源需求的灵活性方面表现出明显的优势。更重要的是,它提供了完全解耦的信号没有交叉对话在双传感功能。像这样的传感器能够监测各种动态人体运动,使其成为下一代人工智能产品的极具前景的候选产品。
    Lightweight and highly compressible materials have received considerable attention in flexible pressure sensing devices. In this study, a series of porous woods (PWs) are produced by chemical removal of lignin and hemicellulose from natural wood by tuning treatment time from 0 to 15 h and extra oxidation through H2O2. The prepared PWs with apparent densities varying from 95.9 to 46.16 mg/cm3 tend to form a wave-shaped interwoven structure with improved compressibility (up to 91.89 % strain under 100 kPa). The sensor assembled from PW with treatment time of 12 h (PW-12) exhibits the optimal piezoresistive-piezoelectric coupling sensing properties. For the piezoresistive properties, it has high stress sensitivity of 15.14 kPa-1, covering a wide linear working pressure range of 0.06-100 kPa. For its piezoelectric potential, PW-12 shows a sensitivity of 0.443 V·kPa-1 with ultralow frequency detection as low as 0.0028 Hz, and good cyclability over 60,000 cycles under 0.41 Hz. The nature-derived all-wood pressure sensor shows obvious superiority in the flexibility for power supply requirement. More importantly, it presents fully decoupled signals without cross-talks in the dual-sensing functionality. Sensor like this is capable of monitoring various dynamic human motions, making it an extremely promising candidate for the next generation artificial intelligence products.
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  • 文章类型: Journal Article
    世界卫生组织(WHO)《2021年全球结核病报告》指出,利福平耐药结核病(RR-TB)仍然是主要的公共卫生威胁。然而,实践中的RR-TB诊断技术有多种限制,包括更长的时间,缺乏敏感性,且检测不到低比例的异质耐药。
    在这里,我们开发了一种基于多重LNA探针的RAP方法(MLP-RAP),用于更灵敏地检测RR-TB的多点突变及其异质抗性。从国家结核病参考实验室收集的126个临床分离株和78个痰样本,中国疾控中心,通过MLP-RAP检测。并行,还进行巢式PCR产物测定的qPCR和Sanger测序以进行比较。
    使用重组质粒,MLP-RAP测定的灵敏度可以达到5拷贝/μl,比qPCR(100拷贝/μl)灵敏20倍。此外,对利福平异质耐药的检测能力为5%。MLP-RAP测定对核酸提取的要求低(煮沸法),并且当置于荧光qPCR仪器中时,反应可以在1小时内完成。临床评价结果表明,MLP-RAP方法可以覆盖516、526、531和533个密码子,具有良好的特异性。经MLP-RAP检测,78份煮沸痰标本中有41份呈阳性,通过巢式PCR产物测定的Sanger测序进一步证实了这一点,相反,qPCR仅能够检测32个样品。与Sanger测序的巢式PCR产物法比较,MLP-RAP检测的特异性和敏感性均为100%.
    MLP-RAP测定可以检测RR-TB感染,具有很高的敏感性和特异性,表明该方法具有应用于一般实验室的快速和灵敏的RR-TB检测的前景。
    UNASSIGNED: The World Health Organization (WHO) Global tuberculosis Report 2021 stated that rifampicin-resistant tuberculosis (RR-TB) remains a major public health threat. However, the in-practice diagnostic techniques for RR-TB have a variety of limitations including longer time, lack of sensitivity, and undetectable low proportion of heterogeneous drug resistance.
    UNASSIGNED: Here we developed a multiplex LNA probe-based RAP method (MLP-RAP) for more sensitive detection of multiple point mutations of the RR-TB and its heteroresistance. A total of 126 clinical isolates and 78 sputum samples collected from the National Tuberculosis Reference Laboratory, China CDC, were tested by MLP-RAP assay. In parallel, qPCR and Sanger sequencing of nested PCR product assay were also performed for comparison.
    UNASSIGNED: The sensitivity of the MLP-RAP assay could reach 5 copies/μl using recombinant plasmids, which is 20 times more sensitive than qPCR (100 copies/μl). In addition, the detection ability of rifampicin heteroresistance was 5%. The MLP-RAP assay had low requirements (boiling method) for nucleic acid extraction and the reaction could be completed within 1 h when placed in a fluorescent qPCR instrument. The result of the clinical evaluation showed that the MLP-RAP method could cover codons 516, 526, 531, and 533 with good specificity. 41 out of 78 boiled sputum samples were detected positive by MLP-RAP assay, which was further confirmed by Sanger sequencing of nested PCR product assay, on the contrary, qPCR was able to detect 32 samples only. Compared with Sanger sequencing of nested PCR product assay, both the specificity and sensitivity of the MLP-RAP assay were 100%.
    UNASSIGNED: MLP-RAP assay can detect RR-TB infection with high sensitivity and specificity, indicating that this assay has the prospect of being applied for rapid and sensitive RR-TB detection in general laboratories where fluorescent qPCR instrument is available.
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  • 文章类型: Journal Article
    背景:聚合酶链反应(PCR)已广泛用于许多病原体检测。然而,PCR技术仍然存在检测时间长、灵敏度不足的问题。重组酶辅助扩增(RAA)是一种功能强大的核酸检测工具,具有较高的灵敏度和扩增效率,但其复杂的探针和无法多重检测阻碍了该技术的进一步应用。
    方法:在本研究中,我们开发并验证了人腺病毒3(HADV3)的多重逆转录重组酶辅助PCR(多重RT-RAP)检测方法,人腺病毒7(HADV7),和人呼吸道合胞病毒(HRSV)在1小时内以人RNaseP蛋白为参考基因监测整个过程。
    结果:使用重组质粒,多重RT-RAP检测HADV3、HADV7和HRSV的灵敏度为每个反应18、3和18个拷贝,分别。多重RT-RAP显示与其他呼吸道病毒没有交叉反应,证明了其良好的特异性。通过多重RT-RAP测试了总共252个临床样本,发现结果与相应的RT-qPCR测定的结果一致。在对选定的阳性标本进行系列稀释测试后,多重RT-RAP的检测灵敏度比相应的RT-qPCR高2至8倍。
    结论:我们得出结论,多重RT-RAP是一种稳健的,快速,高度敏感,和特异性测定,有可能用于筛选低病毒载量的临床样品。
    BACKGROUND: Polymerase chain reaction (PCR) has been widely used for many pathogen detection. However, PCR technology still suffers from long detection time and insufficient sensitivity. Recombinase-aided amplification (RAA) is a powerful nucleic acid detection tool with high sensitivity and amplification efficiency, but its complex probes and inability of multiplex detection hinder the further application of this technology.
    METHODS: In this study, we developed and validated the multiplex reverse transcription recombinase-aided PCR (multiplex RT-RAP) assay for human adenovirus 3 (HADV3), human adenovirus 7 (HADV7), and human respiratory syncytial virus (HRSV) within 1 h with Human RNaseP protein as a reference gene to monitor the whole process.
    RESULTS: Using recombinant plasmids, the sensitivity of multiplex RT-RAP for the detection of HADV3, HADV7, and HRSV was 18, 3, and 18 copies per reaction, respectively. The multiplex RT-RAP showed no cross-reactivity with other respiratory viruses, demonstrating its good specificity. A total of 252 clinical specimens were tested by multiplex RT-RAP and the results were found to be consistent with those of corresponding RT-qPCR assays. After testing serial dilutions of selected positive specimens, the detection sensitivity of multiplex RT-RAP was two to eightfold higher than that of corresponding RT-qPCR.
    CONCLUSIONS: We conclude the multiplex RT-RAP is a robust, rapid, highly sensitive, and specific assay with the potential to be used in the screening of clinical samples with low viral load.
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  • 文章类型: Journal Article
    在动脉粥样硬化中,局部患病区域的慢性炎症过程可能导致活性氧(ROS)的积累。在这项研究中,我们设计了一种高灵敏的H2O2清除纳米仿生系统,负载普罗布考(RPP-PU),更有效地治疗动脉粥样硬化.RPP材料对H2O2具有较高的敏感性,反应灵敏度可从40μmol/L降至10μmol/L,接近病理环境中H2O2的最低浓度水平。RPP-PU延缓了PU的释放并延长了其在体内的持续时间。在载脂蛋白E缺陷(ApoE-/-)小鼠中,RPP-PU有效消除病理性ROS,降低脂质和相关代谢酶的水平,并显著减少血管斑块和纤维的面积。我们的研究表明,清除H2O2的纳米仿生系统可以清除动脉粥样硬化病变中丰富的ROS,从而减少用于治疗动脉粥样硬化的氧化应激,并因此更理想地实现动脉粥样硬化的治疗目标。
    In atherosclerosis, chronic inflammatory processes in local diseased areas may lead to the accumulation of reactive oxygen species (ROS). In this study, we devised a highly sensitive H2O2-scavenging nano-bionic system loaded with probucol (RPP-PU), to treat atherosclerosis more effectively. The RPP material had high sensitivity to H2O2, and the response sensitivity could be reduced from 40 to 10 μmol/L which was close to the lowest concentration of H2O2 levels of the pathological environment. RPP-PU delayed the release and prolonged the duration of PU in vivo. In Apolipoprotein E deficient (ApoE‒/‒) mice, RPP-PU effectively eliminated pathological ROS, reduced the level of lipids and related metabolic enzymes, and significantly decreased the area of vascular plaques and fibers. Our study demonstrated that the H2O2-scavenging nano-bionic system could scavenge the abundant ROS in the atherosclerosis lesion, thereby reducing the oxidative stress for treating atherosclerosis and thus achieve the therapeutic goals with atherosclerosis more desirably.
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