环介导等温扩增技术的应用进展

环介导等温扩增(loop-mediated isothermal amplification,LAMP)是一种新式核酸扩增技术,它依靠一种具有链置换活性的DNA聚合酶与2对特殊设计的引物,在等温条件下即可高效快速地完成扩增反应。相较于传统扩增检测方法,LAMP技术具有特异性强、灵敏度高、操作简单快速等优点,更能在现场快速检测和基层应用中广泛推广,目前LAMP技术已广泛应用于植物病害检测、动物病害检测、食品安全检测等领域。基于此,简要介绍了LAMP技术的基本原理、反应产物的检测方法,重点阐述了LAMP技术的改进与发展,综述了近年来其在科研生产中的应用进展,并对其发展前景进行了展望,以期为LAMP技术的进一步发展提供合理的研究方向。     

环介导等温扩增技术的常见问题分析与研究进展

环介导等温扩增(LAMP)技术是近年发展起来的新型核酸体外等温扩增技术,具有快速、特异、敏感、简便等特点,使该技术在核酸快速检测领域得到了广泛的应用,但LAMP技术也在不断完善与发展中。本文就LAMP扩增产物的污染,引物间非连续配对的假阳性问题的对策,以及在LAMP技术基础上发展起来的其他相关技术进行综述,为该技术在基层的推广应用提供参考。     

等温扩增技术及其结合CRISPR在微生物快速检测中的研究进展

等温扩增技术因其对仪器依赖性低、核酸扩增高效等优势,非常适合于快速检测,已在微生物快速检测领域得到了广泛应用。本文从核酸提取、等温扩增(以环介导等温扩增技术(Loop-mediated isothermal amplification,LAMP)和重组酶聚合酶扩增技术(Recombinase polymerase amplification,RPA)为例)和产物检测角度,就近年来核酸等温扩增技术的发展及其在病原微生物核酸快速检测领域的应用进行综述,并概述了核酸等温扩增技术与CRISPR(Clustered regularly interspaced short palindromic repeats)基因编辑技术相结合的最新研究成果,为这些新兴技术的研究和未来的发展提供新思路。     

快速检测HBV DNA的环状介导等温DNA扩增法

环状介导等温DNA扩增（LAMP）技术是一种新的核酸扩增方法,它能够高特异性、高效、快速地进行核酸的扩增。利用LAMP法检测乙型肝炎病毒（HBV）,能够在等温条件下于1h内将少量的基因拷贝数扩增至10^9,在对65份临床标本的检测中显示了较高的特异性。与现有的PCR技术相比,LAMP法更加简便快速,且在等温条件下进行,不需要复杂的仪器设备,为临床检测乙肝病毒提供了一个快速筒便的新方法。     

环介导等温扩增技术在转基因水稻Bt63检测中的应用

采用环介导等温扩增技术(Loop-Mediated Isothermal Amplification,LAMP)对转基因水稻Bt63进行应用检测。研究表明,环介导等温扩增技术检测转基因水稻Bt63特异性好,并且灵敏度达到0.01%(W/W),具有简便、快速、准确等特点,可以在进出口检验检疫、食品安全监测等领域进行应用。     

环介导等温扩增技术的研究进展

环介导等温扩增(loop-mediated isothermal amplification,LAMP)是一种新式核酸扩增技术,它依靠一种具有链置换活性的DNA聚合酶和2对特殊设计的引物,不需要反复的温度循环和昂贵的仪器设备,在等温条件下即可高效快速地完成扩增反应,目前已广泛应用于细菌、病毒、寄生虫等病原体的检测,及动物胚胎性别的鉴定。该文总结了LAMP技术的基本原理、相对于传统核酸检测技术的优点、产物的检测方法及其临床应用,最后指出LAMP目前存在的不足以及采取的相应措施,并对其发展前景进行了展望。     

环介导等温扩增核酸技术及其在食品安全检测领域的应用

环介导等温扩增(Loop-mediated isothermal amplification,简称LAMP)是利用能识别靶序列上6个位点的4个特殊设计的引物和一种具有链置换活性的DNA聚合酶,在恒温条件下,特异、高效、快速地扩增核酸的新技术。该技术在1h内扩增效率可达到109-1010个数量级,扩增产物是一系列反向重复的靶序列构成的茎环结构和多环花椰菜样结构的DNA片段混合物,电泳后在凝胶上显现出由不同大小的区带组成的阶梯式图谱。近年来LAMP技术以其特异性强、等温灵敏、操作简单、产物易检测等优点已经应用于食品安全检测领域的多个方面。     

多重环介导等温扩增技术的研究进展

环介导等温扩增技术(LAMP)是一种新型的核酸放大扩增技术。但是,当前的LAMP技术多是在同一个体系中对单一目标物进行检测,限制了其工作效率的发挥。多重LAMP技术就是在同一反应体系中加入多组针对不同靶基因的特异性引物,从而实现对多种目标基因同时进行扩增。我们针对多重LAMP的优缺点,对其在病毒、细菌、寄生虫以及性别筛选等领域的应用做简要综述。     

环介导等温扩增技术的改进及其在现场检测平台上的应用进展

环介导等温扩增技术(Loop-mediated isothermal amplification,LAMP)是一种新型核酸体外扩增技术,具有等温、快速、高特异性和灵敏度、产物检测简便等特点。自建立以来,该技术被广泛应用于病原检测、动物胚胎的性别鉴定、转基因食品检测和肿瘤的基因检测等领域并得到不断改进和完善,主要体现在特异性提高,反应速度加快,样本处理简化,检测方法特异化,多重扩增得以实现。由于LAMP满足现场检测的诸多要求,当前基于LAMP技术的现场检测平台正向着微型化、集成化、自动化方向发展,并呈现出与芯片实验室和数字扩增等技术联合应用的趋势。本文综述了近年来LAMP技术的改进及其在现场检测平台上的应用进展。     

环介导等温扩增技术作为基因快速诊断方法的研究与展望

环介导等温扩增技术(LAMP)是一种新型分子诊断技术,具有灵敏度高、反应快速等优点,且仅利用简单的恒温水浴设备即可实现对特异性靶基因的高效扩增,在分子诊断领域具有巨大潜力。近几年来LAMP技术在分子诊断领域的研究非常广泛,涉及细菌、病毒、寄生虫的检测及食品转基因成分检验等诸多方面,受到越来越多学者认可,有研究者认为其可作为PCR的替补技术应用于临床基因诊断工作中。然而由于目前该技术缺乏规范化且成熟配套的上下游技术措施,如快速简单的核酸抽提技术、完善的应用体系和方案、防止交叉污染的有效措施及小型便携的检测设备等,因此在现场即时诊断领域的推广应用较为缓慢。为发挥利用LAMP技术在快速基因诊断方面的优越性,科研人员针对LAMP技术开发应用存在的问题开展了大量研究工作。立足于临床基因快速诊断的技术需求,对影响LAMP技术应用的核酸物质的快速抽提、LAMP检测体系的优化完善、结果判定、假阳性结果的规避、结果确诊及现场诊断一体化平台设计应用等领域的发展现状、需要攻克的难点及未来的应用展望进行综述,以期为未来的研究和开发指明方向,促进LAMP技术的不断完善及临床推广应用。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.