LAMP

  • 文章类型: Journal Article
    环介导等温扩增(LAMP)技术是基于PCR的方法的绝佳替代方法,因为它很快,易于使用,具有高灵敏度和特异性,无需昂贵的仪器。然而,LAMP的局限性之一是难以实现在单个管中同时检测多个目标,作为允许这种方法依赖于含有特定靶序列的荧光探针的方法,使它们的适应和测定的优化复杂化。这里,我们总结了基于序列特异性检测的多重LAMP检测的不同方法,用技术的示意图来说明,并根据结果的实时检测和量化评估其实际应用,一目了然地可视化结果的可能性,反应组分的预先稳定,促进即时护理使用,扩增的特异性靶标的最大数量,以及该技术在临床样本中的验证。各种LAMP多路复用方法在其操作条件和机制方面不同。每种方法都有其优点和缺点,它们之间的选择将取决于特定的应用兴趣。
    The loop-mediated isothermal amplification (LAMP) technique is a great alternative to PCR-based methods, as it is fast, easy to use and works with high sensitivity and specificity without the need for expensive instruments. However, one of the limitations of LAMP is difficulty in achieving the simultaneous detection of several targets in a single tube, as the methodologies that allow this rely on fluorogenic probes containing specific target sequences, complicating their adaptation and the optimization of assays. Here, we summarize different methods for the development of multiplex LAMP assays based on sequence-specific detection, illustrated with a schematic representation of the technique, and evaluate their practical application based on the real-time detection and quantification of results, the possibility to visualize the results at a glance, the prior stabilization of reaction components, promoting the point-of-care use, the maximum number of specific targets amplified, and the validation of the technique in clinical samples. The various LAMP multiplexing methodologies differ in their operating conditions and mechanism. Each methodology has its advantages and disadvantages, and the choice among them will depend on specific application interests.
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  • 文章类型: Systematic Review
    我们荟萃分析了快速诊断测试(试纸)和环介导等温扩增(LAMP)方法检测志贺氏菌的诊断准确性。我们搜索了MEDLINE,Embase,WebofScience和GoogleScholar从成立到2023年的研究报告了志贺氏菌试纸和LAMP测试与培养或聚合酶链反应(PCR)相比的性能。我们的搜索确定了2618项研究,其中14项符合系统审查的纳入标准。包括4056项测试(来自12个国家)的10项研究被纳入荟萃分析。总体合并的敏感性和特异性分别为98%(95%CI:94-100)和97%(95%CI:92-99),分别。试纸的集合敏感性和特异性分别为95%和98%,分别。相比之下,LAMP显示出更高的合并敏感度(100%)和诊断优势比(431752),但特异性相似(97%)。LAMP和量油尺测试表现出良好的性能,这表明它们可能有助于诊断志贺氏菌病。
    We meta-analyzed the diagnostic accuracy of rapid diagnostic tests (dipsticks) and loop-mediated isothermal amplification (LAMP) method to detect Shigella species. We searched MEDLINE, Embase, Web of Science and Google Scholar from inception to 2023 for studies reporting on the performance of Shigella dipstick and LAMP tests compared with culture or polymerase chain reaction (PCR). Our search identified 2618 studies, of which fourteen met the inclusion criteria for the systematic review. Ten studies covering 4056 tests (from twelve countries) were included in the meta-analysis. The overall pooled sensitivity and specificity were 98% (95% CI: 94-100) and 97% (95% CI: 92-99), respectively. Pooled sensitivity and specificity of dipsticks were 95% and 98%, respectively. In contrast, LAMP showed higher pooled sensitivity (100%) and diagnostic odds ratio (431752), but similar specificity (97%). LAMP and dipstick tests exhibited promising performance, suggesting that they could be useful for assisting in the diagnosis of shigellosis.
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  • 文章类型: Journal Article
    人类真菌感染,特别是由念珠菌和曲霉引起的,已成为主要的公共卫生负担。常规诊断的长周转时间和差的灵敏度是更快诊断人类真菌病原体的主要障碍。
    为了克服这些问题,已经开发了基于分子的诊断。它们提供增强的灵敏度,但需要复杂的基础设施,熟练的人力,并且仍然昂贵。在这种情况下,环介导等温扩增(LAMP)分析代表了一种有希望的替代方法,可以促进视觉读出。然而,为了根除真菌感染,必须准确检测所有形式的真菌。因此,迫切需要快速的替代测试方法,准确,便于广泛采用。因此,本研究的目的是使用科学数据库进行荟萃分析,以评估LAMP在根据PRISMA指南检测一组人类真菌病原体中的诊断效率。PubMed,谷歌学者,科学直接,Scopus,BioRxiv,还有MedRxiv.
    从各种关于真菌诊断的研究报告来看,只有9篇文章被鉴定为符合基于LAMP的诊断标准.通过这个荟萃分析,发现大多数研究是在中国和日本进行的,痰和血液是最常用的LAMP检测标本。收集的数据强调ITS基因和基于荧光的检测被列为最常用的靶标和方法。荟萃分析的合并敏感性值介于0.71和1.0之间,森林地块和SROC(受试者工作特征摘要)曲线显示出合并特异性值介于0.13和1.0之间,置信区间为95%,分别。符合条件的研究的准确率和准确率大多在70%至100%和68%至100%之间变化,分别。进行了基于QUADAS-2(诊断准确性研究质量评估)的偏倚和适用性的质量评估,该评估描述了低偏倚风险和适用性问题。一起,LAMP技术可以被认为是当前诊断的可行替代方案,考虑到高真菌负担,可以在低资源地区进行快速测试。
    UNASSIGNED: Human fungal infections particularly caused by Candida and Aspergillus have emerged as major public health burden. Long turnaround time and poor sensitivity of the conventional diagnostics are the major impediments for faster diagnosis of human fungal pathogens.
    UNASSIGNED: To overcome these issues, molecular-based diagnostics have been developed. They offer enhanced sensitivity but require sophisticated infrastructure, skilled manpower, and remained expensive. In that context, loop-mediated isothermal amplification (LAMP) assay represents a promising alternative that facilitates visual read outs. However, to eradicate fungal infections, all forms of fungi must be accurately detected. Thus, a need for alternative testing methodologies is imperative that should be rapid, accurate and facilitate widespread adoption. Therefore, the aim of the present study is to conduct a meta-analysis to assess the diagnostic efficiency of LAMP in the detection of a panel of human fungal pathogens following PRISMA guidelines using scientific databases viz. PubMed, Google Scholar, Science Direct, Scopus, BioRxiv, and MedRxiv.
    UNASSIGNED: From various studies reported on the diagnosis of fungi, only 9 articles were identified as eligible to meet the criteria of LAMP based diagnosis. Through this meta-analysis, it was found that most of the studies were conducted in China and Japan with sputum and blood as the most common specimens to be used for LAMP assay. The collected data underlined that ITS gene and fluorescence-based detections ranked as the most used target and method. The pooled sensitivity values of meta-analysis ranged between 0.71 and 1.0 and forest plot and SROC (summary receiver operating characteristic) curve revealed a pooled specificity values between 0.13 and 1.0 with the confidence interval of 95%, respectively. The accuracy and precision rates of eligible studies mostly varied between 70 to 100% and 68 to 100%, respectively. A quality assessment based on QUADAS-2 (Quality Assessment of Diagnostic Accuracy Studies) of bias and applicability was conducted which depicted low risk of bias and applicability concerns. Together, LAMP technology could be considered as a feasible alternative to current diagnostics considering high fungal burden for rapid testing in low resource regions.
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  • 文章类型: Journal Article
    结核病(TB)仍然是影响全球三分之一人口的致命疾病。常规诊断的长周转时间和差的灵敏度是快速诊断分枝杆菌属以防止耐药性的主要障碍。为了克服这些问题,分子诊断已经开发出来。它们提供增强的灵敏度,但需要复杂的基础设施,熟练的人力和仍然昂贵。
    在这种情况下,环介导等温扩增(LAMP)分析,世卫组织在2016年推荐用于结核病诊断,听起来是一种有希望的替代方案,有助于视觉读出。因此,本研究的目的是进行荟萃分析,以评估LAMP检测一组分枝杆菌的诊断效率。遵循使用科学数据库的PRISMA指南。1600项研究报道了分枝杆菌的诊断。,入选的30篇文章符合基于LAMP的诊断标准.
    研究发现,大多数研究都是在印度等高疾病负担国家进行的,泰国,和日本,痰是最常用的LAMP检测标本。此外,IS6110基因和基于荧光的检测分别是最常用的目标和方法。准确率和准确率大多在79.2%至99.3%和73.9%至100%之间。分别。最后,进行了基于QUADAS-2偏倚和适用性的质量评估.
    LAMP技术可以被认为是当前诊断的可行替代方案,考虑到在低资源区域进行快速测试的高负担。
    UNASSIGNED: Tuberculosis (TB) remains a deadly disease affecting one-third population globally. Long turnaround time and poor sensitivity of the conventional diagnostics are the major impediments for faster diagnosis of Mycobacterial spp to prevent drug resistance. To overcome these issues, molecular diagnostics have been developed. They offer enhanced sensitivity but require sophisticated infrastructure, skilled manpower and remain expensive.
    UNASSIGNED: In that context, loop-mediated isothermal amplification (LAMP) assay, recommended by the WHO in 2016 for TB diagnosis, sounds as a promising alternative that facilitates visual read outs. Therefore, the aim of the present study is to conduct a meta-analysis to assess the diagnostic efficiency of LAMP for the detection of a panel of Mycobacterium spp. following PRISMA guidelines using scientific databases. From 1600 studies reported on the diagnosis of Mycobacterium spp., a selection of 30 articles were identified as eligible to meet the criteria of LAMP based diagnosis.
    UNASSIGNED: It was found that most of the studies were conducted in high disease burden nations such as India, Thailand, and Japan with sputum as the most common specimen to be used for LAMP assay. Furthermore, IS6110 gene and fluorescence-based detections ranked as the most used target and method respectively. The accuracy and precision rates mostly varied between 79.2% to 99.3% and 73.9% to 100%, respectively. Lastly, a quality assessment based on QUADAS-2 of bias and applicability was conducted.
    UNASSIGNED: LAMP technology could be considered as a feasible alternative to current diagnostics considering high burden for rapid testing in low resource regions.
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  • 文章类型: Retraction of Publication
    背景:植物病原体在本领域通常通过它们可能引起的典型疾病症状来鉴定。有效的早期检测和识别病原体是采取有效的管理措施以减少或防止其传播以减轻疾病的负面影响的基本程序。在这里,在这篇综述中提出并讨论了早期检测病原体的传统和创新方法。此外,他们强调了当前技术的主要优点和局限性。
    结果:用于植物病原体鉴定的传统诊断技术通常集中在DNA上,RNA(当分子方法),和蛋白质或肽(当血清学方法)的病原体。基于主要酶联免疫吸附测定(ELISA)的血清学方法是用于病原体检测的最常用方法,因为它们具有高通量潜力和低成本。这种技术对于无症状感染阶段的许多病原体检测不是特别可靠和足够灵敏。对于实验室中不可培养的病原体,基于核酸的技术是一致的病原体检测或鉴定的最佳选择。横向流动系统是创新的工具,即使在现场条件下也能快速准确地获得结果,但是他们有需要克服的敏感性问题。在上一代便携式热循环仪上进行的PCR测定可以在原位提供快速检测结果。
    结论:便携式仪器的出现可以加快病原体检测,降低商业成本,并有可能彻底改变植物病理学。这篇综述提供了有关有效检测不同植物病原体的当前方法和程序的信息。本文受版权保护。保留所有权利。
    Next-generation methods for early disease detection in crops. A review. (https://doi.org/10.1002/ps.7471) Daniela Trippa, Riccardo Scalenghe, Marcos Fernando Basso, Stefano Panno, Salvatore Davino, Chiara Morone, Antonio Giovino, Safa Oufensou, Nicola Luchi, Sanaz Yousefi and Federico Martinelli. The above article from Pest Management Science, published online on 24 March 2023 in Wiley Online Library (http://wileyonlinelibrary.com) has been withdrawn by agreement among the Journal Editors-in-Chief, Dr Stephen Duke and Professor Yidong Wu, and John Wiley & Sons Ltd on behalf of the Society of Chemical Industry. The withdrawal has been agreed because of extensive revisions after acceptance of the paper. The new study is being resubmitted to the journal for formal review. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
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  • 文章类型: Journal Article
    字数:4645,数字数:4,表格数:12019年12月COVID-19疫情引发急性呼吸道疾病全球大流行,随着突变株的毒力和确诊病例数的增加,这对全球公共卫生造成了巨大威胁。因此,为了快速控制SARS-CoV-2的传播,迫切需要准确诊断COVID-19。作为一种新的分子生物学技术,环介导等温扩增(LAMP)操作方便,速度,成本低,灵敏度和特异性高。在过去的两年里,COVID-19的猖獗和病毒株的不断变异对病原体的快速检测提出了更高的要求。与常规RT-PCR和实时RT-PCR方法相比,基因分型RT-LAMP方法和LAMP加肽核酸(PNA)探针检测方法已被开发出来,以正确鉴定SARS-CoV-2变体,这也是为什么LAMP技术备受关注的原因。LAMP检测技术结合侧流分析,微流控技术和其他传感技术可以通过核酸扩增有效地增强信号,并有助于更快地给出所产生的输出,更方便和用户友好的方式。目前,LAMP在SARS-CoV-2的检测中起着重要作用。
    The number of words: 4645, the number of figures: 4, the number of tables: 1The outbreak of COVID-19 in December 2019 caused a global pandemic of acute respiratory disease, and with the increasing virulence of mutant strains and the number of confirmed cases, this has resulted in a tremendous threat to global public health. Therefore, an accurate diagnosis of COVID-19 is urgently needed for rapid control of SARS-CoV-2 transmission. As a new molecular biology technology, loop-mediated isothermal amplification (LAMP) has the advantages of convenient operation, speed, low cost and high sensitivity and specificity. In the past two years, rampant COVID-19 and the continuous variation in the virus strains have demanded higher requirements for the rapid detection of pathogens. Compared with conventional RT-PCR and real-time RT-PCR methods, genotyping RT-LAMP method and LAMP plus peptide nucleic acid (PNA) probe detection methods have been developed to correctly identified SARS-CoV-2 variants, which is also why LAMP technology has attracted much attention. LAMP detection technology combined with lateral flow assay, microfluidic technology and other sensing technologies can effectively enhance signals by nucleic acid amplification and help to give the resulting output in a faster, more convenient and user-friendly way. At present, LAMP plays an important role in the detection of SARS-CoV-2.
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  • 文章类型: Journal Article
    微生物沾染惹起的食源性疾病一向是人类极其存眷的事宜。因此,对这些问题的研究从未停止过。随着微生物扩增技术的发展,越来越多的检测方法进入了我们的视野。然而,传统的检测技术或多或少存在缺陷,例如复杂的操作,精度低,灵敏度低,长时间检测,等等。因此,更方便,准确,并且需要对微生物进行灵敏的测量。等温扩增技术是包含上述优点的替代方法之一。本文主要总结了环介导等温扩增(LAMP)和滚环扩增(RCA)的原理。同时,介绍了LAMP和RCA在食品微生物检测中的应用。
    Food-borne diseases caused by microbial contamination have always been a matter of great concern to human beings. Hence, the research on these problems has never stopped. With the development of microorganism amplification technology, more and more detection methods have come into our vision. However, traditional detection technologies presents more or less drawbacks, such as complicated operation, low accuracy, low sensitivity, long-time detection, and so on. Therefore, more convenient, accurate, and sensitive measurement for the microorganism are needed. Isothermal amplification technology is one of the alternative approach containing the above mentioned advantages. This work mainly summarizes the principles of loop-mediated isothermal amplification (LAMP) and rolling circle amplification (RCA) which belong to isothermal amplification. Meanwhile, the application of LAMP and RCA in food microorganism detection is introduced.
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  • 文章类型: Journal Article
    世界上已经进行了超过60亿次的COVID-19测试。SARS-CoV-2(严重急性呼吸系统综合症冠状病毒-2)病毒和相应的人类抗体的测试不仅对于医疗机构诊断和治疗感染至关重要,也是国际航班等重大半正常经济和社会活动的先决条件,在办公室离线工作和学习,进入商场,体育和社交活动。准确性,灵敏度,特异性,时间的结果和成本的每个测试是这些测试的基本参数,甚至最小的改进,其中任何一个可能有显著的影响,在世界许多国家的生活。我们描述,对COVID-19检测方法进行了分析和比较,同时在22个表中表示它们的参数。此外,我们比较了一些FDA批准的检测试剂盒的测试性能和一些刚刚在科学文献中描述的非FDA批准的方法的临床性能.RT-PCR仍然是检测该病毒的金标准,但是迫切需要更便宜的替代品,更快,护理方法很明显。解释了那些最终可能会被开发来满足这一需求的方法,讨论,定量比较。这篇综述具有生物分析化学的前瞻性,但对于有兴趣了解和改进COVID-19检测的更广泛的读者来说,这可能会很有趣,帮助最终离开过去的COVID-19大流行。
    More than six billion tests for COVID-19 has been already performed in the world. The testing for SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus-2) virus and corresponding human antibodies is essential not only for diagnostics and treatment of the infection by medical institutions, but also as a pre-requisite for major semi-normal economic and social activities such as international flights, off line work and study in offices, access to malls, sport and social events. Accuracy, sensitivity, specificity, time to results and cost per test are essential parameters of those tests and even minimal improvement in any of them may have noticeable impact on life in the many countries of the world. We described, analyzed and compared methods of COVID-19 detection, while representing their parameters in 22 tables. Also, we compared test performance of some FDA approved test kits with clinical performance of some non-FDA approved methods just described in scientific literature. RT-PCR still remains a golden standard in detection of the virus, but a pressing need for alternative less expensive, more rapid, point of care methods is evident. Those methods that may eventually get developed to satisfy this need are explained, discussed, quantitatively compared. The review has a bioanalytical chemistry prospective, but it may be interesting for a broader circle of readers who are interested in understanding and improvement of COVID-19 testing, helping eventually to leave COVID-19 pandemic in the past.
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  • 文章类型: Journal Article
    一种新酵母的识别,耳念珠菌,2009年在东亚,以及它在全球的迅速传播,提醒人们多药耐药真菌病原体构成的威胁。由于无法获得准确的测试,C.auris可能长时间未被识别。实验室社区通过在2014年至2021年初之间发布35种新的或修订的诊断方法来应对C.auris的挑战。商业部门还修改了现有的诊断设备。这些C.auris诊断测试从传统的基于文化的差异和选择性培养基中运行,生化同化,和快速的蛋白质图谱,以及培养无关的基于DNA的诊断。我们提供了这些发展的概述,特别是具有验证数据的测试,这些数据随后被用于通用.我们共享实验室开发的工作流程,以处理纽约都会区爆发的37,000个C.auris监测样本和5,000个C.auris分离株。我们的预览涵盖了基于微流体的新设备和诊断方法,光学,和纳米技术。前线实验室需要快速,便宜,稳定,和易于实施的测试,以提高C.auris诊断,监视,患者隔离,入院筛查,和环境控制。迫切需要的是用于推定C.auris鉴定的侧向流测定或类似装置。所有实验室都将受益于允许快速抗真菌药敏测试的设备,包括检测赋予耐药性的突变。希望,多重测试小组即将实现C.auris测试与其他医疗保健相关感染的持续监测的协同作用。C.auris基因组分析在疫情调查中具有被证明的作用,和诊断实验室需要快速访问区域和国家基因组分析网络。
    The recognition of a new yeast, Candida auris, in 2009 in East Asia, and its rapid global spread, was a reminder of the threats posed by multidrug-resistant fungal pathogens. C. auris had likely remained unrecognized for a long time as accurate tests were not available. The laboratory community responded to the C. auris challenge by publishing 35 new or revised diagnostic methods between 2014 and early 2021. The commercial sector also modified existing diagnostic devices. These C. auris diagnostic tests run the gamut from traditional culture-based differential and selective media, biochemical assimilations, and rapid protein profiles, as well as culture-independent DNA-based diagnostics. We provide an overview of these developments, especially the tests with validation data that were subsequently adopted for common use. We share a workflow developed in our laboratory to process over 37,000 C. auris surveillance samples and 5,000 C. auris isolates from the outbreak in the New York metropolitan area. Our preview covers new devices and diagnostic approaches on the horizon based on microfluidics, optics, and nanotechnology. Frontline laboratories need rapid, cheap, stable, and easy-to-implement tests to improve C. auris diagnosis, surveillance, patient isolation, admission screening, and environmental control. Among the urgent needs is a lateral flow assay or similar device for presumptive C. auris identification. All laboratories will benefit from devices that allow rapid antifungal susceptibility testing, including detection of mutations conferring drug resistance. Hopefully, multiplex test panels are on the horizon for synergy of C. auris testing with ongoing surveillance of other healthcare-associated infections. C. auris genome analysis has a proven role for outbreak investigations, and diagnostic laboratories need quick access to regional and national genome analysis networks.
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  • 文章类型: Journal Article
    BACKGROUND: Schistosomiasis is a parasitic disease caused by hematodes of genus Schistosoma. This review evaluated the available nucleic acid amplification techniques for diagnosing S. mansoni infections in humans, intermediate host snails, and presumed rodent reservoirs.
    METHODS: Sensitivity, specificity, diagnostic odds ratio (DOR), and 95% CI were calculated based on available literature. The potential of PCR, nPCR, PCR-ELISA, qPCR, and LAMP was compared for diagnosing S. mansoni infections.
    RESULTS: A total of 546 published records were identified. Quality assessment by QUADAS-2 revealed an uncertain risk in most studies, and 21 references were included in the final. For human samples, the four nucleic acid amplification techniques showed an overall sensitivity of 89.79% (95% CI: 83.92%-93.67%), specificity of 87.70% (95% CI: 72.60%-95.05%), and DOR of 37.73 (95% CI: 21.79-65.33). LAMP showed the highest sensitivity, followed by PCR-ELISA, PCR, and qPCR, while this order was almost reversed for specificity; qPCR had the highest AUC. For rodent samples, qPCR showed modest sensitivity (68.75%, 95% CI: 43.32%-86.36%) and high specificity (92.45%, 95% CI: 19.94%-99.83%). For snail samples, PCR and nPCR assays showed high sensitivity of 90.06% (95% CI: 84.39%-93.82%) and specificity of 85.51% (95% CI: 54.39%-96.69%).
    CONCLUSIONS: Nucleic acid amplification techniques had high diagnostic potential for identifying S. mansoni infections in humans.
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