TwistDx推出世界上首个以重组酶聚合酶扩增技术为基础的DNA扩增试剂盒

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TwistDx Launches World's First DNA Amplification Kit Based on Recombinase Polymerase Amplification (RPA) 7/17/2009 DNA扩增RPA取代PCR? ——TwistDx全球发售基于重组聚合酶扩增的第一代DNA扩增试剂盒 CAMBRIDGE, England--(BUSINESS WIRE)--TwistDx, a developer of point-of-use nucleic acid diagnostics, announced today the launch of its TwistAmpTM product range, the world’s first DNA amplification products based on Recombinase Polymerase Amplification (RPA). Concurrently, TwistDx also launched the TwistaTM portable, real-time fluorometer, an easy-to-use RPA reader. RPA is a proprietary technique, employing recombinase enzymes, used to amplify nucleic acids in any biological sample, with high analytical sensitivity and specificity. Each of the four currently available TwistAmpTM kits (outlined below) offers scientists the ability to detect DNA molecules in a mixed sample within 10-15 minutes, less than half the time of most PCR systems. Moreover, RPA operates at low, constant temperatures and does not even require initial melting of the sample DNA, opening the way to broader access to end-users without reliance on expensive thermocyclers. RPA has also been shown to be highly resistant to crude samples in comparison to PCR suggesting applications in on-the-spot field testing requiring no nucleic acid extraction. The TwistaTM portable real-time fluorometer is a customised platform containing proprietary fluorescent probe systems developed by the company to accurately monitor RPA reactions. Although seamlessly integrated with the TwistaTM device, TwistAmpTM kits can be used in combination with various, widely used assay platforms, such as conventional analysis tools (e.g. on agarose gels), dipsticks, as well as broadly available fluorescence plate readers and real-time PCR machines. “Our TwistAmpTM kit offers researchers a flexible and cost-effective solution to achieve DNA amplification in significantly less time. With laboratory analysis a critical but often time-consuming aspect of biomedical research, we believe these new research tools provide our customers with a viable alternative to PCR, which although very effective is costly to access and time consuming,” said Dr. Niall Armes, CEO of TwistDx. “In particular, real-time PCR devices are very expensive; a laboratory could purchase multiple TwistaTM devices for the same price. Field researchers may also be able to easily perform their measurements in a mobile setting as TwistAmpTM enzyme and nucleotide mixes are provided in a freeze-dried form that demonstrates good stability at ambient temperatures. The TwistaTM device will be available with portable battery pack and standalone software.” The TwistAmpTM product range can be ordered through the TwistDx website (www.twistdx.co.uk). Details of each product can be found below: TwistAmp TM Basic The TwistAmpTM Basic kit contains all enzymes and reagents necessary for the amplification of DNA. Primers and the template must be supplied by the user. Amplification success is typically assessed by an end-point method, such as gel electrophoresis. Amplified material can also be purified and used for down-stream applications (e.g. subcloning). TwistAmpTM exo The TwistAmpTM exo kit is recommended for users who want to combine the TwistDx amplification technology with the company’s proprietary fluorescent exo GapProbeTm in a homogenous format. In addition to the basic components, it contains a powerful nuclease (Exonuclease III) which processes exo GapProbeTm during the amplification reaction and generates a real-time read-out. The presence of Exonuclease III reduces the final overall yield of amplified material at endpoint and so is not suitable for analysis on gels, however it is the preferred system for generating strong fluorescence signal kinetics in the RPA system. TwistAmpTM nfo The TwistAmpTM nfo kit is designed for users who want to detect the success of their amplification reaction by means of end-point ‘sandwich-assays’, such as lateral-flow (LF) strips. Besides the basic amplification reagents, it includes a nuclease (nfo) that can generate new polymerase extension substrates within suitable LF GapProbeTm. The amplified material can then be used in instrument–free detection formats. The TwistAmpTM nfo kit can also be used for fluorescence monitoring using fluorescent exo GapProbeTm as an alternative to the TwistAmpTM exo kit as nfo processes these probes (often with slightly slower kinetics) and will in addition permit product to be analysed by gels at endpoint. TwistAmpTM fpg The TwistAmpTM fpg kit is tailored for users who want to combine the TwistDx amplification technology with an alternative TwistDx reporter probe system, fluorescent fpg GapProbeTM, in a homogenous format. These probes have less design constraints than exo GapProbeTm, although kinetics of fluorescence accumulation may be slower. In addition to the basic components it contains a powerful nuclease (fpg) that processes fpg GapProbeTm during the amplification reaction itself and generate a real-time read-out. The presence of fpg will not reduce the final overall yield of amplified material, in contrast to the use of Exonuclease III in the TwistAmpTM exo kit, allowing end point gel analysis, but the kinetics of probe cutting can be slower than that with the exo system. TwistAmpTM RT The TwistAmpTM RT kit will contain all enzymes and reagents necessary for the amplification of RNA – all that has to be supplied by the user are the primers and the template. TwistaTM portable real-time fluorometer The TwistaTM portable real-time fluorometer is a customised device developed for monitoring RPA reactions containing TwistDx’s fluorescent probe systems, exo GapProbeTm, fpg GapProbeTm, or intercalating dyes should they be employed. The TwistaTM contains a heated incubation chamber for a strip of 8 tubes perfectly designed for the RPA reagents supplied in the TwistAmpTM kits, and can monitor fluorescence in real-time in two colour channels with exquisite sensitivity. The TwistaTM has a small footprint and can be rendered entirely portable with the addition of a rechargeable battery pack. Data can be analysed by PC-link, or alternatively the device can function as a standalone unit capable of storing and running user-defined methods to call samples positive and negative for any characterised assay. About TwistDx TwistDx, Ltd., based in Cambridge, UK, is a privately-held diagnostic company that is developing novel tests based on breakthrough DNA amplification technology. These tests retain the highest possible levels of specificity and sensitivity, while adding extraordinary speed and portability compared to current DNA diagnostics in use. Ultimately the company believes this new generation of tests will provide gold standard DNA testing globally in point of care and field settings. TwistDx’s revolutionary technology, Recombinase Polymerase Amplification (RPA), is positioned to become the DNA-based diagnostic platform of the future, with wide ranging applications in a number of fields, including medical diagnostics, veterinary medicine, industrial applications, agriculture, public health and biosecurity and environmental sensing. More information about TwistDx is available at www.twistdx.co.uk

英国剑桥——（商业电报）——开发使用点核酸诊断的TwistDx公司宣布推出TwistAmpTM产品系列，其是世界上首个以重组酶聚合酶扩增（RPA）技术为基础的DNA扩增产品。同时，TwistDx也推出了TwistaTM便携式实时定量计，其是易于使用的RPA读取器。RPA是一种专利技术，其利用重组酶，用于扩增任何一种生物标本的核酸，具有很高的分析敏感性和特异性。

目前推出了4种TwistAmpTM试剂盒，每一种TwistAmpTM试剂盒（列于下面）使科学家能够在10-15分钟内检测DNA分子，较多数PCR系统的一半时间都短。而且，RPA在低、常温运转，不需要样本DNA的初始变性，给使用者提供了更广泛的途径，而无需依靠昂贵的热循环仪。RPA可以使用粗提样本，与之相对，PCR说明中需要抽提核酸。

TwistaTM便携式实时定量计是一可以定制的平台，包含有公司开发的合适的荧光探针系统，以精确测定RPA反应。TwistAmpTM试剂盒可以与TwistaTM仪器无缝连接，也可以与多种广泛使用的检测平台，如传统分析工具（如琼脂糖胶）、分析仪、荧光读取器以及实时定量PCR仪联合使用。

TwistDx公司CEO Niall Armes说：“我们的TwistAmpTM试剂盒为研究者提供了（方法上）灵活、价钱上划算的在十分短的时间内取得DNA扩增结果的方法。当实验室分析生物医学研究的某个关键方面，但是耗费时间，我们相信这些新研究工具为客户提供了可行的PCR取代方法，尽管取得结果花费大。尤其是，实时PCR设备很昂贵，一个实验室可以同样的价格购买多个TwistaTM设备。由于TwistAmpTM的酶和核苷酸混合物是以冻干形式提供，其对室温稳定，实地研究者可以很容易地操作测量。TwistaTM设备可以便携电池组和独立软件获得。”

TwistAmpTM产品系列可以通过TwistDx网站订购。每种产品的细节如下：

TwistAmp TM Basic TwistAmpTM Basic试剂盒包含所有的扩增DNA必须的酶和试剂。用户需要提供引物和模板。扩增成功由终点方法评估，如琼脂糖电泳。扩增材料可以被纯化在后续试验使用（如亚克隆）。

TwistAmpTM exo TwistAmpTM exo试剂盒推荐给联合使用TwistDx扩增技术和该公司的专利荧光外切核酸酶GapProbeTm的用户。除了基本组成部分外，还包括强大的核酸酶（核酸外切酶III），其在扩增反应期间行使外切酶活性，进行实时读出。核酸外切酶III减少终点时扩增材料的最终总体产率，所以不适于用琼脂糖胶检测，然而其是RPA系统中产生强莹光信号的首选信号。

TwistAmpTM nfo TwistAmpTM nfo试剂盒为通过终点“三明治测定”（如侧向流动条）检测扩增反应成功者设计的。除基本扩增试剂外，包括核酸外切酶（nfo），其在合适的LF GapProbeTm内可以产生新的多聚酶延伸底物。扩增材料接下来可被用于无器械检测。TwistAmpTM nfo试剂盒也可以被应用于荧光监测，通过使用荧光核酸外切酶GapProbeTm作为TwistAmpTM exo试剂盒替代品，处理这些探针（常常具有弱的动力学），允许产物使用琼脂糖胶进行终点检测。

TwistAmpTM fpg TwistAmpTM fpg试剂盒是特为想把TwistDx扩增技术与TwistDx报告探针系统和荧光fpg GapProbeTM二者择一相结合者设计的。尽管荧光的动态堆积可能缓慢，这些探针较核酸外切酶GapProbeTm有较少的设计参数限制。除了基本组成部分，还包括强大的核酸酶（fpg），其在扩增反应期间处理fpg GapProbeTm本身，产生实时读出。与TwistAmpTM exo 试剂盒使用核酸外切酶III相对比，fpg不减少最终扩增材料的总体产率，允许终点琼脂糖电泳分析，但是探针切下的动力学较exo系统缓慢。

TwistAmpTM RT TwistAmpTM RT试剂盒包含扩增RNA所必须的酶和试剂——用户需要提供的仅仅是引物和模板。

TwistaTM便携实时荧光计 TwistaTM便携实时荧光计是一种用户定制的设备，用以监测包含TwistDx’s荧光探针系统、核酸外切酶GapProbeTm、fpg GapProbeTm或应该使用增补燃料在内的RPA反应。TwistaTM包含有一个加热孵育室，可以孵育TwistaAmpTM试剂盒所提供的为RPA试剂设计的8个试管，还可以在2个颜色通道非常精确监测实时荧光。TwistaTM占用面积小，当使用可再充电的蓄电池时可以成为一便携设备。可以通过连接个人计算机分析数据，该设备也可以作为一个单独的装置存储、运行用户定义的调用阳性和阴性样本测定的方法。

TwistDx TwistDx公司，以英国剑桥为基础，是一个私有诊断公司，其以DNA扩增技术突破为基础开发新的实验设备。这些实验设备具有高水平的特异性和敏感性，与目前使用的DNA诊断（设备）相比具有速度快和便携性特点。该公司相信其新一代的产品将提供DNA检测的金标准。TwistDx公司的革命性技术——重组酶多聚酶扩增，定位于成为将来DNA诊断平台，可在多个领域有广泛的应用，包括医学诊断、兽医学、工业应用、农业公共卫生与生物安全以及环境测知。关于TwistDx公司的更多信息可以访问网址www.twistdx.co.uk。