detection method

检测方法
  • 文章类型: Journal Article
    猪deltacoronavirus(PDCoV)是一种新发现的猪肠道致病性冠状病毒,具有单链正义RNA基因组和包膜。PDCoV感染不同年龄的猪,并导致新生仔猪急性腹泻和呕吐。在严重的情况下,感染导致脱水,疲惫,和病态仔猪的死亡,给养猪场带来巨大的经济损失。PDCoV感染的临床症状与其他猪肠道病毒非常相似。尽管不进行测试很难区分这些病毒感染,监测PDCoV非常重要,因为它可以在人群中传播。最常用的检测PDCoV的方法是qPCR,这是耗时的,需要熟练的人员和设备。许多农场无法满足检测所需的条件。因此,有必要建立一种更快速、更方便的检测PDCoV的方法。
    通过将RPA(重组酶聚合酶等温扩增)与CRISPR/Cas13a相结合,建立一种快速便捷的PDCoV检测方法。
    设计了PDCoV的特异性RPA引物和crRNA,样品中的核酸用RPA扩增。进行荧光CRISPR/Cas13a检测。我们使用qPCR作为对照方法评估了RPA-CRISPR/Cas13a测定的灵敏度和特异性。
    CRISPR/Cas13a辅助检测在90分钟内完成。PDCoV的最低检测限为5.7×101拷贝/μL。特异性分析显示该测定不与其它三种猪肠道病毒发生交叉反应。
    RPA-CRISPR/Cas13a方法具有灵敏度高的优点,特异性强,快速反应,和容易获得的结果,并可用于PDCoV的检测。
    UNASSIGNED: Porcine deltacoronavirus (PDCoV) is a newly discovered porcine intestinal pathogenic coronavirus with a single-stranded positive-sense RNA genome and an envelope. PDCoV infects pigs of different ages and causes acute diarrhea and vomiting in newborn piglets. In severe cases, infection leads to dehydration, exhaustion, and death in sick piglets, entailing great economic losses on pig farms. The clinical symptoms of PDCoV infection are very similar to those of other porcine enteroviruses. Although it is difficult to distinguish these viral infections without testing, monitoring PDCoV is very important because it can spread in populations. The most commonly used methods for the detection of PDCoV is qPCR, which is time-consuming and require skilled personnel and equipment. Many farms cannot meet the conditions required for detection. Therefore, it is necessary to establish a faster and more convenient method for detecting PDCoV.
    UNASSIGNED: To establish a rapid and convenient detection method for PDCoV by combining RPA (Recombinase Polymerase Isothermal Amplification) with CRISPR/Cas13a.
    UNASSIGNED: Specific RPA primers and crRNA for PDCoV were designed, and the nucleic acids in the samples were amplified with RPA. Fluorescent CRISPR/Cas13a detection was performed. We evaluated the sensitivity and specificity of the RPA-CRISPR/Cas13a assay using qPCR as the control method.
    UNASSIGNED: CRISPR/Cas13a-assisted detection was completed within 90 min. The minimum detection limit of PDCoV was 5.7 × 101 copies/μL. A specificity analysis showed that the assay did not cross-react with three other porcine enteroviruses.
    UNASSIGNED: The RPA-CRISPR/Cas13a method has the advantages of high sensitivity, strong specificity, fast response, and readily accessible results, and can be used for the detection of PDCoV.
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  • 文章类型: Journal Article
    众多具有优异的光捕获和光热转换能力的纳米材料已广泛应用于各个领域,如光热诊断和治疗,微量物质检测,和光学成像。尽管近年来已经建立了利用纳米材料的光热效应的光热检测方法,关于它们在食品安全检测中的应用的评论很少。在这里,光热转换机制的最新进展,光热转换效率计算,综述了光热纳米材料的制备方法。特别是,全面论述了光热纳米材料在各种食品危害分析中的应用以及新建立的光热检测方法。此外,讨论了基于光热纳米材料的检测方法的发展和未来的发展趋势,这为研究者提出更有效的,敏感,和准确的检测方法。
    Numerous nanomaterials endowed with outstanding light harvesting and photothermal conversion abilities have been extensively applied in various fields, such as photothermal diagnosis and therapy, trace substance detection, and optical imaging. Although photothermal detection methods have been established utilizing the photothermal effect of nanomaterials in recent years, there is a scarcity of reviews regarding their application in food safety detection. Herein, the recent advancements in the photothermal conversion mechanism, photothermal conversion efficiency calculation, and preparation method of photothermal nanomaterials were reviewed. In particular, the application of photothermal nanomaterials in various food hazard analyses and the newly established photothermal detection methods were comprehensively discussed. Moreover, the development and promising future trends of photothermal nanomaterial-based detection methods were discussed, which provide a reference for researchers to propose more effective, sensitive, and accurate detection methods.
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  • 文章类型: Journal Article
    蔬菜病害的自动检测和识别可以提高蔬菜品质,增加利润。温室种植的蔬菜疾病的图像通常具有复杂的背景,各种各样的疾病,和微妙的症状差异。以前的研究已经努力准确地确定病变位置和量化感染程度,导致整体识别率较低。为了应对验证数据集不足以及检测和识别率低带来的挑战,本研究利用寿光的地理优势,被誉为“蔬菜镇”,\"建立自建蔬菜基地进行数据采集和验证实验。集中于患有各种疾病的广泛的水果和蔬菜作物,我们进行了温室病害图像的现场采集,编制了一个大规模的数据集,并介绍了时空融合注意力网络(STFAN)。STFAN整合了蔬菜病害发生的多源信息,增强模型的弹性。此外,我们提出了多层编码器-解码器特征融合网络(MEDFFN)来抵消深度卷积块中的特征消失,辅以边界结构损失函数,以指导模型获取更详细,更准确的边界信息。通过设计从多个源提取高分辨率特征表示的检测和识别模型,实现了疾病的精确检测和识别。本研究为蔬菜病害的整体防控提供了技术支持,从而推进智慧农业。结果表明,在我们自建的VDGE数据集上,与YOLOv7-tiny相比,YOLOv8n,和YOLOv9,提出的模型(多源信息融合方法,用于蔬菜病害检测,MIFV)使MAP提高了3.43%,3.02%,和2.15%,分别,展示显著的性能优势。MIFV模型参数为39.07M,计算复杂度为108.92GFLOPS,突出突出的实时性和检测精度相比主流算法。这项研究表明,提出的MIFV模型可以以更低的成本快速准确地检测和识别温室环境中的蔬菜疾病。
    Automated detection and identification of vegetable diseases can enhance vegetable quality and increase profits. Images of greenhouse-grown vegetable diseases often feature complex backgrounds, a diverse array of diseases, and subtle symptomatic differences. Previous studies have grappled with accurately pinpointing lesion positions and quantifying infection degrees, resulting in overall low recognition rates. To tackle the challenges posed by insufficient validation datasets and low detection and recognition rates, this study capitalizes on the geographical advantage of Shouguang, renowned as the \"Vegetable Town,\" to establish a self-built vegetable base for data collection and validation experiments. Concentrating on a broad spectrum of fruit and vegetable crops afflicted with various diseases, we conducted on-site collection of greenhouse disease images, compiled a large-scale dataset, and introduced the Space-Time Fusion Attention Network (STFAN). STFAN integrates multi-source information on vegetable disease occurrences, bolstering the model\'s resilience. Additionally, we proposed the Multilayer Encoder-Decoder Feature Fusion Network (MEDFFN) to counteract feature disappearance in deep convolutional blocks, complemented by the Boundary Structure Loss function to guide the model in acquiring more detailed and accurate boundary information. By devising a detection and recognition model that extracts high-resolution feature representations from multiple sources, precise disease detection and identification were achieved. This study offers technical backing for the holistic prevention and control of vegetable diseases, thereby advancing smart agriculture. Results indicate that, on our self-built VDGE dataset, compared to YOLOv7-tiny, YOLOv8n, and YOLOv9, the proposed model (Multisource Information Fusion Method for Vegetable Disease Detection, MIFV) has improved mAP by 3.43%, 3.02%, and 2.15%, respectively, showcasing significant performance advantages. The MIFV model parameters stand at 39.07 M, with a computational complexity of 108.92 GFLOPS, highlighting outstanding real-time performance and detection accuracy compared to mainstream algorithms. This research suggests that the proposed MIFV model can swiftly and accurately detect and identify vegetable diseases in greenhouse environments at a reduced cost.
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  • 文章类型: Journal Article
    解决当前铁路轨道异物检测技术的局限性,其缺点是实时性能不足和检测小物体的准确性降低,本文介绍了一种创新的基于视觉的感知方法,利用深度学习的力量。这种方法的核心是利用复杂的轨道检测方法构建铁路边界模型,以及增强的UNet语义分割网络,以实现对不同轨道类别的自主分割。通过采用等间隔除法和逐行遍历,精确提取临界航迹特征点,通过最小二乘法推导出轨道线性方程,从而建立了准确的铁路边界模型。我们从四个方面对YOLOv5s检测模型进行了优化:将SE注意机制纳入颈部网络层,以增强模型的特征提取能力,添加预测层以提高小物体的检测性能,提出了一种线性尺寸缩放方法来获得合适的锚盒,利用Inner-IoU细化边界回归损失函数,从而提高边界框的定位精度。我们使用自构建的图像数据集对铁路轨道异物入侵进行了检测准确性验证。结果表明,所提出的语义分割模型实现了91.8%的MIoU,比以前的模型提高了3.9%,有效地分割铁路轨道。此外,优化后的检测模型可以有效地检测异物对轨道的侵入,与原始YOLOv5s模型相比,减少了漏报和误报,平均精度提高了7.4%(IoU=0.5)。该模型在涉及小对象的场景中表现出强大的泛化能力。该方法对铁路轨道异物入侵检测的深度学习技术进行了有效探索,适合在复杂环境中使用,确保铁路线的运行安全。
    Addressing the limitations of current railway track foreign object detection techniques, which suffer from inadequate real-time performance and diminished accuracy in detecting small objects, this paper introduces an innovative vision-based perception methodology harnessing the power of deep learning. Central to this approach is the construction of a railway boundary model utilizing a sophisticated track detection method, along with an enhanced UNet semantic segmentation network to achieve autonomous segmentation of diverse track categories. By employing equal interval division and row-by-row traversal, critical track feature points are precisely extracted, and the track linear equation is derived through the least squares method, thus establishing an accurate railway boundary model. We optimized the YOLOv5s detection model in four aspects: incorporating the SE attention mechanism into the Neck network layer to enhance the model\'s feature extraction capabilities, adding a prediction layer to improve the detection performance for small objects, proposing a linear size scaling method to obtain suitable anchor boxes, and utilizing Inner-IoU to refine the boundary regression loss function, thereby increasing the positioning accuracy of the bounding boxes. We conducted a detection accuracy validation for railway track foreign object intrusion using a self-constructed image dataset. The results indicate that the proposed semantic segmentation model achieved an MIoU of 91.8%, representing a 3.9% improvement over the previous model, effectively segmenting railway tracks. Additionally, the optimized detection model could effectively detect foreign object intrusions on the tracks, reducing missed and false alarms and achieving a 7.4% increase in the mean average precision (IoU = 0.5) compared to the original YOLOv5s model. The model exhibits strong generalization capabilities in scenarios involving small objects. This proposed approach represents an effective exploration of deep learning techniques for railway track foreign object intrusion detection, suitable for use in complex environments to ensure the operational safety of rail lines.
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  • 文章类型: Journal Article
    间变性淋巴瘤激酶(ALK)重排的存在定义了非小细胞肺癌(NSCLC)的分子亚型。ALK抑制剂(ALKIs)在ALK阳性晚期NSCLC患者中具有显著的临床获益;因此,准确选择,快速,和适当的ALK测试方法,以筛选适合抗ALK治疗的患者。近年来,ALKIs的开发和临床应用取得了很大进展,以及我们对获得性耐药机制的理解。同时,新的ALK辅助诊断平台已被开发并应用于临床实践。尽管许多研究表明,这些平台之间的一致性很高,在测试过程中不断出现新的问题。为患者带来最大利益,首先选择合适的测试方法,然后制定,优化,并根据检测人群和标本类型遵守标准化的检测流程。随着临床实践数据的不断积累,ALK检测质量控制的经验,以及多中心研究的结果,更新的专家共识是必要的。参与讨论和制定本指南的专家具有丰富的理论背景和临床检验经验,这确保了本指南中提供的信息的实用价值。
    The presence of anaplastic lymphoma kinase (ALK) rearrangement defines a molecular subtype of non-small cell lung cancer (NSCLC). ALK inhibitors (ALKIs) confer significant clinical benefits in patients with ALK-positive advanced NSCLC; therefore, it is of great clinical significance to select accurate, rapid, and appropriate ALK testing methods to screen for patients who are suitable for anti-ALK treatment. In recent years, great progress has been made in the development and clinical application of ALKIs, as well as in our understanding of acquired drug resistance mechanisms. Meanwhile, new ALK companion diagnostic platforms have been developed and applied in clinical practice. Although many studies have shown that there is a high rate of concordance among these platforms, new problems continue to appear during testing. To maximize the benefit for patients, accurate testing results can be obtained by first selecting the appropriate testing method and then formulating, optimizing, and complying with the standardized testing process in accordance with the testing population and specimen types. With the ongoing accumulation of clinical practice data, experience from quality control of ALK testing, and results from multicenter research, an updated expert consensus is necessary. The experts who participated in the discussion and development of this guideline have a rich background in theoretical and clinical testing experience, which ensures the practical value of the information presented in this guideline.
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  • 文章类型: Journal Article
    猫冠状病毒(FCoV),猫泛白细胞减少症病毒(FPV),猫白血病病毒(FeLV)在中国普遍存在,并严重威胁猫的健康。这些病毒引起类似的表现和病理损害。快速准确的诊断取决于实验室的检测。本研究旨在建立一种可靠、快速、准确检测FCoV的方法,FPV,和FeLV,以便做出明确的诊断,并采取有效措施预防和控制病毒感染。
    我们设计了三对特异性引物和探针,用于检测FCoV5'非翻译区,FPV病毒蛋白2和FeLVpol基因。产生重组质粒构建体用作标准质粒构建体。最佳反应条件,包括引物和探针浓度,反应循环,和退火温度,在优化试验的基础上获得。成功建立了一步三螺旋实时逆转录-定量聚合酶链反应(RT-qPCR)同时检测FCoV,FPV,和FeLV。特异性,灵敏度,并分析了试验的可重复性,并通过检测1175份临床样本验证了其适用性。
    一步三联RT-qPCR仅对FCoV的检测具有高度特异性,FPV,和FeLV;它具有高灵敏度,p-FCoV的检出限为139.904、143.099和152.079拷贝/反应,p-FPV,和p-FeLV标准质粒构建体,分别,它具有可靠的可重复性,测定中的变异系数为0.06%-0.87%。总共对1175个临床样本进行了FCoV检查,FPV,和FeLV使用三重RT-qPCR,还有FCoV,FPV,FeLV阳性率为18.47%,19.91%,和47.57%,分别。一步法三组分RT-qPCR的临床敏感性和特异性分别为93.07%和97.99%,分别。
    我们开发了一种快速可靠的一步法三重RT-qPCR方法,用于检测FCoV,FPV,和FeLV,可以用作临床监测和诊断的诊断工具。
    UNASSIGNED: Feline coronavirus (FCoV), feline panleukopenia virus (FPV), and feline leukemia virus (FeLV) are prevalent throughout China and significantly threaten cat health. These viruses cause similar manifestations and pathological damage. Rapid and accurate diagnosis depends on detection in the laboratory. This study aimed to establish a reliable and rapid method for accurate detection of FCoV, FPV, and FeLV so that a definite diagnosis can be made and effective measures can be taken to prevent and control viral infection.
    UNASSIGNED: We designed three pairs of specific primers and probes for the detection of FCoV 5\' untranslated region, FPV viral protein 2, and FeLV pol genes. Recombinant plasmid constructs were generated for use as standard plasmid constructs. Optimal reaction conditions, including primer and probe concentrations, reaction cycles, and annealing temperatures, were obtained on the basis of optimization tests. One-step triplex real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was successfully established to simultaneously detect FCoV, FPV, and FeLV. The specificity, sensitivity, and repeatability of the assay were analyzed, and its applicability was validated by testing 1175 clinical samples.
    UNASSIGNED: One-step triplex RT-qPCR had a high degree of specificity only for the detection of FCoV, FPV, and FeLV; it had high sensitivity with limits of detection of 139.904, 143.099, and 152.079 copies/reaction for p-FCoV, p-FPV, and p-FeLV standard plasmid constructs, respectively, and it had reliable repeatability with 0.06%-0.87% intra-assay coefficients of variations. A total of 1175 clinical samples were examined for FCoV, FPV, and FeLV using triplex RT-qPCR, and the FCoV, FPV, and FeLV positivity rates were 18.47%, 19.91%, and 47.57%, respectively. The clinical sensitivity and specificity of one-step triplex RT-qPCR were 93.07% and 97.99%, respectively.
    UNASSIGNED: We developed a rapid and reliable one-step triplex RT-qPCR method for the detection of FCoV, FPV, and FeLV, which could be used as a diagnostic tool for clinical monitoring and diagnosis.
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  • 文章类型: Journal Article
    路基损伤具有隐蔽性强、难以检测的特点。出于这个原因,冲击成像方法已用于道路基层的检测。本文系统地讨论了采集点的设置,励磁方式和数据处理方法。通过在公路路面基层灌浆养护前后检测中的应用,结果表明该方法简便、准确。检测结果可以以二维图像形式显示,并且易于在道路维护中使用。该方法可用于识别和定位路面基层的损伤,判断路面基层结构的均匀性。还可用于评价灌浆修复后内部损伤的有效性。
    The damage of road base course has the characteristics of strong concealment and difficulty in detecting. For this reason, the impact imaging method has been used for detection of road base course. This paper discussed systematically collection points setting, excitation mode and data processing method. Through the application in testing for highway pavement base before and after grouting maintenance, the results show that the method is simple and accurate. The detection results can be displayed in a two-dimensional image form and it is easy to be used in road maintenance. This method can be used to identify and locate the damages of the pavement base, to judge the uniformity of the pavement base structure. It can also be used to evaluate the effectiveness of internal damage after grouting repairing.
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  • 文章类型: Journal Article
    膀胱癌(BC)是全球第十大最常见的恶性肿瘤,由于其高发病率,提出了重大的临床和社会挑战,快速发展,频繁复发。目前,膀胱镜检查和尿细胞学检查是已建立的BC诊断方法。然而,其疗效受到侵入性和低敏感性的限制。因此,开发高度特异性的生物标志物和有效的非侵入性检测策略对于实现精确和及时的BC诊断势在必行。以及促进最佳肿瘤治疗和改善预后。microRNAs(miRNAs),短非编码RNA分子跨越约20-25个核苷酸,与多种致癌途径的调节有关。显著改变的miRNA形成强大的功能调节网络,其对BC的肿瘤发生和进展产生显著影响。来自血液的异常miRNAs的研究,尿液,或胞外囊泡表明它们在BC中作为诊断生物标志物和预后指标的潜在作用,使miRNA能够监测疾病的进展并预测疾病的复发。同时,这项以miRNA为中心的研究是一种潜在的治疗药物,为BC的治疗提供了一种新的方法.本文综述了miRNAs在肿瘤发生和发展中的生物学作用。并系统地总结了作为BC的诊断和预后生物标志物以及治疗靶标的潜力。此外,我们评估了该领域实验室技术的进展,并讨论了前景。
    Bladder cancer (BC) is the tenth most prevalent malignancy globally, presenting significant clinical and societal challenges because of its high incidence, rapid progression, and frequent recurrence. Presently, cystoscopy and urine cytology serve as the established diagnostic methods for BC. However, their efficacy is limited by their invasive nature and low sensitivity. Therefore, the development of highly specific biomarkers and effective non-invasive detection strategies is imperative for achieving a precise and timely diagnosis of BC, as well as for facilitating an optimal tumor treatment and an improved prognosis. microRNAs (miRNAs), short noncoding RNA molecules spanning around 20-25 nucleotides, are implicated in the regulation of diverse carcinogenic pathways. Substantially altered miRNAs form robust functional regulatory networks that exert a notable influence on the tumorigenesis and progression of BC. Investigations into aberrant miRNAs derived from blood, urine, or extracellular vesicles indicate their potential roles as diagnostic biomarkers and prognostic indicators in BC, enabling miRNAs to monitor the progression and predict the recurrence of the disease. Simultaneously, the investigation centered on miRNA as a potential therapeutic agent presents a novel approach for the treatment of BC. This review comprehensively analyzes biological roles of miRNAs in tumorigenesis and progression, and systematically summarizes their potential as diagnostic and prognostic biomarkers, as well as therapeutic targets for BC. Additionally, we evaluate the progress made in laboratory techniques within this field and discuss the prospects.
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  • 文章类型: Journal Article
    Levan,β-(2,6)-连接的果糖聚合物,表现出不同的特性,赋予多功能性,使其成为各种工业应用的备受追捧的生物聚合物。Levan可以由各种微生物使用蔗糖生产,食品工业副产品和农业废物。微生物果聚糖代表了用于商业规模果聚糖生产的最有效的成本效益方法。本研究通过了解果聚糖的生物合成来回顾果聚糖生产的优化,理化性质和发酵过程。此外,介绍和讨论了遗传和蛋白质工程以提高其产量以及新兴的检测方法。所有这些综合研究都可以作为优化果聚糖生产并扩大其在各个行业的应用的强大工具。
    Levan, a β-(2,6)-linked fructose polymer, exhibits diverse properties that impart versatility, rendering it a highly sought-after biopolymer with various industrial applications. Levan can be produced by various microorganisms using sucrose, food industry byproducts and agricultural wastes. Microbial levan represents the most potent cost-effective process for commercial-scale levan production. This study reviews the optimization of levan production by understanding its biosynthesis, physicochemical properties and the fermentation process. In addition, genetic and protein engineering for its increased production and emerging methods for its detection are introduced and discussed. All of these comprehensive studies could serve as powerful tools to optimize levan production and broaden its applications across various industries.
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  • 文章类型: Journal Article
    pH和Cu2+离子浓度的变化与阿尔茨海默病和癌症等疾病有关。快速检测pH和Cu2+离子对于公共卫生和环境问题至关重要。半salamo型探针(E)-2-羟基-1-萘醛O-(2-(氨基氧基)乙基)肟(NSS)表现出大量的双功能性能,感应pH变化和Cu2+离子。探头NSS上的单激发和双发射特性使其与众不同。探针NSS表现出pH依赖性激发态分子内质子转移(ESIPT),并且其光学性质根据pH环境而变化。探针NSS通过改变激发态分子内质子转移(ESIPT)机制的“关-关-关”来检测pH从2到11的变化,展示迅速,可逆,选择性反应。此外,发光salamo类萘基探针NSS可以与Cu2+离子配位,以0.84ppb(13.2nM)的检出限实现识别Cu2离子的高选择性和灵敏度探针NSS可以检测实际水样中的Cu2离子,例如自来水和黄河水。装载有探针NSS的测试条能够快速准确地检测水样中的Cu2+离子。因此,多功能salamo型探针NSS为开发高灵敏度和快速响应的双模式pH计以及Cu2传感奠定了基础。
    pH and Cu2+ ion concentration changes are linked to disorders like Alzheimer\'s and cancer. Rapid detection of pH and Cu2+ ions is critical for public health and environmental concerns. The semi-salamo-type probe (E)-2-hydroxy-1-naphthaldehyde O-(2-(aminooxy)ethyl) oxime (NSS) demonstrated substantial dual-functional performance, sensing pH change and Cu2+ ions. A single excitation and double emission characteristic on the probe NSS made it distinctive. Probe NSS exhibits pH-dependent excited state intramolecular proton transfer (ESIPT), and its optical properties vary based on the pH environment. Probe NSS detects pH changes from 2 to 11 by changing the \"off-on-off\" of the excited state intra-molecular proton transfer (ESIPT) mechanism, exhibiting rapid, reversible, and selective responses. In addition, the luminescent salamo-like naphthalene-based probe NSS can coordinate with Cu2+ ions, achieving great selectivity and sensitivity to identify Cu2+ ions with a detection limit of 0.84 ppb (13.2 nM) Probe NSS can detect Cu2+ ions in actual water samples such as tap water and yellow river water. The test strip loaded with probe NSS enabled quick and accurate detection of Cu2+ ions in water samples. Consequently, the versatile salamo-type probe NSS lays the foundation for developing high sensitivity and fast-response dual-mode pH meters as well as Cu2+ sensing.
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