Detection method

检测方法
  • 文章类型: Journal Article
    一种新的创新方法,MICA军团菌,允许在2天内自动计数生活水样中的嗜肺军团菌,每个测试部分的检测限为2CFU。在这里,我们表明它在7到15天内给出了与法国标准方法NFT90-431获得的结果相同的结果。
    A new innovative method, MICA Legionella, allows for the automatic enumeration of Legionella pneumophila in domestic water samples in 2 days, with a detection limit of 2 CFU per test portion. Here we show that it gives equivalent results to those obtained by the French standard method NF T90-431 in 7 to 15 days.
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  • 文章类型: Journal Article
    Triatomine虫子是克氏锥虫的载体,美洲大陆查加斯病的病因。这里,我们已经测试了环介导等温扩增(LAMP)测试,用于直接检测Triatomainfestans粪便中的Cruzi。这种寄生虫在美国南锥体的主要媒介。分析评估显示,用与每个离散分型单位(I-VI)相对应的克鲁兹T.Cruzi菌株的DNA人工接种的triatomine粪便样品的阳性结果,每个反应的灵敏度高达一个寄生虫。相反,用rangeli锥虫和其他系统发育相关和无关生物体的DNA测试时,反应产生阴性结果。在真实野外条件下(从城市家庭)捕获的三叶草中,并使用参考显微镜技术定义为T.Cruzi阳性或阴性,LAMP测试达到了100%的一致性。我们的结果表明,这种LAMP反应表现出优异的分析特异性和灵敏度,没有来自粪便基质的干扰,因为所有反应都是在没有纯化步骤的情况下进行的。这种简单的分子诊断技术可以很容易地由病媒控制机构在现场条件下使用。
    Triatomine bugs are vectors of Trypanosoma cruzi, the etiologic agent of Chagas disease in the American continent. Here, we have tested a loop-mediated isothermal amplification (LAMP) test for a direct detection of T. cruzi in feces of Triatoma infestans, the main vector of this parasite in the Southern Cone of America. The analytical evaluation showed positive results with samples of triatomine feces artificially inoculated with DNA from strains of T. cruzi corresponding to each Discrete Typing Units (I-VI), with a sensitivity of up to one parasite per reaction. Conversely, the reaction yielded negative results when tested with DNA from Trypanosoma rangeli and other phylogenetically related and unrelated organisms. In triatomines captured under real field conditions (from urban households), and defined as positive or negative for T. cruzi using the reference microscopy technique, the LAMP test achieved a concordance of 100 %. Our results demonstrate that this LAMP reaction exhibits excellent analytical specificity and sensitivity without interference from the fecal matrix, since all the reactions were conducted without purification steps. This simple molecular diagnostic technique can be easily used by vector control agencies under field conditions.
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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
    包括水稻在内的20多种转基因粮食作物已在许多国家获得批准。转基因水稻及其衍生产品尚未在印度获得批准,因此在印度被视为未经授权的转基因生物。因此,重要的是要跟踪是否含有大米食品,市场上可用的是无转基因的。
    进行了一项试点研究,以检查30个包装米粒和以大米为原料的加工食品样品的转基因状态,使用针对花椰菜花叶病毒35S启动子(P-35S)的聚合酶链反应(PCR)测定法,茉莉碱合成酶终止子(T-nos),膦丝菌素-N-乙酰转移酶(pat)和cry1Ac基因,这可以涵盖所有全球批准的转基因水稻事件的筛查。
    根据结果,测试的样本均未发现P-35S阳性,T-nos,pat和cry1Ac。
    在测试的样品中未检测到未经授权的转基因大米成分。可以进一步进行此类研究,以测试从各个谷类作物的转基因事件获得批准的国家进口的食品中除大米以外的谷物的转基因成分。作为监管要求的一部分。
    UNASSIGNED: More than 20 genetically modified (GM) food crops including rice have been approved in many countries. GM rice and derived products have not yet been approved in India so they are considered as unauthorized genetically modified organisms (GMOs) in the country. Therefore it is important to track whether the rice containing food items, available in the marketplace are GMO-free.
    UNASSIGNED: A pilot study was conducted to check the GM status of 30 samples of packed rice grains and processed food products with rice as an ingredient, using polymerase chain reaction (PCR) assays targeting Cauliflower Mosaic Virus 35 S promoter (P-35 S), nopaline synthase terminator (T-nos), phosphinothricin-N-acetyltransferase (pat) and cry1Ac gene, which could cover screening for all the globally approved GM rice events.
    UNASSIGNED: Based on the results, none of the samples tested were found positive for P-35 S, T-nos, pat and cry1Ac.
    UNASSIGNED: The unauthorized presence of GM rice ingredients was not detected in the samples tested. Such studies may further be conducted for the testing of GM ingredients derived from cereals other than rice in the food products imported from the country where GM events of respective cereal crop are approved, as a part of regulatory requirement.
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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 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 provides a comprehensive analysis of the biological roles of miRNAs in tumorigenesis and progression, and systematically summarizes the potential as diagnosis and prognosis 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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  • 文章类型: Journal Article
    蚕豆真花叶病毒(BBTMV)感染蚕豆和豌豆,降低产量。由于BBTMV通过蚕豆传播,许多国家已经实施了防止受感染种子分发的法规。目前,酶联免疫吸附测定(ELISA)通常用于检测BBTMV。而基于PCR的方法由于其灵敏度和速度而优选用于种子病毒检测。尚未报道BBTMV特异性PCR检测方法。目前存在一种通用的检测方法,该方法利用逆转录PCR(RT-PCR)来检测Comovirus属,BBTMV所属。然而,物种鉴定需要序列分析。为了解决这个限制,我们使用新设计的BBTMV特异性引物开发并验证了RT-PCR检测方法。使用这些引物的RT-PCR和实时RT-PCR比ELISA敏感约5×105-106倍,比以前报道的RT-PCR方法敏感100-1,000倍。使用这些引物使用RT-PCR和实时RT-PCR,当每克蚕豆种子存在超过3.0×105个拷贝时,我们可以以相同的灵敏度检测BBTMV。我们新开发的检测方法可以高灵敏度和快速地检测BBTMV。
    Broad bean true mosaic virus (BBTMV) infects broad beans and peas, reducing yield. As BBTMV is transmitted through broad beans, many countries have implemented regulations to prevent the distribution of infected seeds. Currently, enzyme-linked immunosorbent assay (ELISA) is commonly used to detect BBTMV. While the PCR-based method is preferred for seed virus detection due to its sensitivity and speed. A BBTMV-specific PCR detection method has not yet been reported. A universal detection method currently exists that utilizes reverse transcription PCR (RT-PCR) for the Comovirus genus, to which BBTMV belongs. However, sequence analysis is required for species identification. To address this limitation, we developed and verified RT-PCR detection methods using newly designed BBTMV-specific primers. RT-PCR and real-time RT-PCR with these primers were approximately 5 × 105-106 times more sensitive than ELISA and 100-1000 times more sensitive than previously reported RT-PCR methods. Using RT-PCR and real-time RT-PCR employing these primers, we could detect BBTMV with same sensitivity when more than 3.0 × 105 copies were present per gram of broad bean seeds. Our newly developed detection methods can test for BBTMV with high sensitivity and speed.
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  • 文章类型: Journal Article
    猪呼吸道冠状病毒(PRCoV),猪繁殖与呼吸综合征病毒(PRRSV),猪流感病毒(SIV),伪狂犬病病毒(PRV)是引起猪呼吸道疾病的重要病毒。病猪表现出类似的临床症状,如发烧,咳嗽,流鼻涕,呼吸困难,这使得很难在现场准确鉴别诊断这些疾病。在这项研究中,四重一步逆转录实时定量PCR(RT-qPCR)检测PRCoV,PRRSV,SIV,PRV成立。该测定法显示出强特异性,高灵敏度,和良好的重复性。它只能检测到PRCoV,PRRSV,SIV,和PRV,没有与TGEV的交叉反应,PEDV,PRoV,ASFV,FMDV,PCV2,PDCoV,CSFVPRCoV的检测限(LOD),PRRSV,SIV,PRV为129.594、133.205、139.791和136.600拷贝/反应,分别。测定内和测定间变异系数(CV)范围为0.29%至1.89%。建立的四重RT-qPCR用于检测4909个临床标本,这些都是在广西收集的,中国,从2022年7月到2023年9月。PRCoV,PRRSV,SIV,PRV的阳性率为1.36%,10.17%,4.87%,和0.84%,分别。此外,先前报道的RT-qPCR也用于测试这些样本,这些方法之间的一致性高于99.43%。建立的四重RT-qPCR可以同时准确检测这四种猪呼吸道病毒,为临床诊断提供准确可靠的检测技术。
    Porcine respiratory coronavirus (PRCoV), porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV), and pseudorabies virus (PRV) are significant viruses causing respiratory diseases in pigs. Sick pigs exhibit similar clinical symptoms such as fever, cough, runny nose, and dyspnea, making it very difficult to accurately differentially diagnose these diseases on site. In this study, a quadruplex one-step reverse-transcription real-time quantitative PCR (RT-qPCR) for the detection of PRCoV, PRRSV, SIV, and PRV was established. The assay showed strong specificity, high sensitivity, and good repeatability. It could detect only PRCoV, PRRSV, SIV, and PRV, without cross-reactions with TGEV, PEDV, PRoV, ASFV, FMDV, PCV2, PDCoV, and CSFV. The limits of detection (LODs) for PRCoV, PRRSV, SIV, and PRV were 129.594, 133.205, 139.791, and 136.600 copies/reaction, respectively. The intra-assay and inter-assay coefficients of variation (CVs) ranged from 0.29% to 1.89%. The established quadruplex RT-qPCR was used to test 4909 clinical specimens, which were collected in Guangxi Province, China, from July 2022 to September 2023. PRCoV, PRRSV, SIV, and PRV showed positivity rates of 1.36%, 10.17%, 4.87%, and 0.84%, respectively. In addition, the previously reported RT-qPCR was also used to test these specimens, and the agreement between these methods was higher than 99.43%. The established quadruplex RT-qPCR can accurately detect these four porcine respiratory viruses simultaneously, providing an accurate and reliable detection technique for clinical diagnosis.
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