酶活性和动力学行为是生物化学和催化化学研究中非常重要的内容。本文综述了微量量热技术在酶催化反应中的最新研究和应用进展,重点介绍了等温滴定量热和流动微量量热技术,并讨论了微量量热技术在计算酶催化反应动力学参数方面的优势。

Determination of enzyme kinetics and activity are the most important analytical procedures in biochemistry. The present review deals with the research progress of the microcalorimetry of enzyme-catalyzed reaction. The more recent applications of calorimetric techniques such as isothermal titration cal-orimetry and flow calorimetry to the study of enzyme kinetics ,as well as the advantages on using calorime-tric techniques in the determination of kinetic parameters of enzymes ,is also discussed here.

【摘要】目的建立实时荧光环介导等温扩增快速检测结核分枝杆菌的方法。方法针对结核分枝杆菌特异序列建立实时荧光环介导等温扩增技术；对其特异性和敏感性进行评估；通过敏感性和特异性等指标验证其稳定性。结果建立的实时荧光环介导等温扩增法对结核和非结核分枝杆菌标准菌株检测的敏感性和特异性均为100％，灵敏度可达102个菌／mL；痰标本结核分歧杆菌检测的敏感性为94．64％（53，56），荧光定量聚合酶链式反应方法检测的敏感性为92．86％（52／56）；检测的特异性为96．55％（28，29），荧光定量聚合酶链式反应方法检测的特异性为96．55％（28／29）。结论建立实时荧光环介导等温扩增检测结核分枝杆菌的方法，敏感性和特异性强，灵敏度高，稳定性好，可用于痰液中结核分枝杆菌的快速检测。

Objective To establish a rapid method of real-time fluorescence loop-mediated isothermal amplification (LAMP) for detection of Mycobacterium tuberculosis. Methods According to the specific sequence of IS6110 in Mycobacterium tuberculosis, 6 primers of loop-mediated isothermal amplification were designed and synthesized. With the ESE-Quant tube scanner platform, a real-time fluorescence loop-mediated isothermal amplification was established. 14 strains of non-tuberculous mycobacteria standard strains and 14 strains of Mycobacterium tuberculosis isolated from cases were used to evaluate specificity and sensitivity of the method. Sputum samples were collected from 56 positive cases for Mycobacterium tuberculosis and 29 negative cases for Mycobacterium tuberculosis. The stability of the method was validated by the sensitivity and specificity compared to the real-time fluorescent PCR method. Results Real-time fluorescence loop-mediated isothermal amplification method was established, the sp

环介导等温扩增(LAMP)技术是一种新型核酸扩增技术，该技术在60～65℃恒温条件1 h内，把特异性靶序列扩增到109水平，具有高特异性、高敏感性、简单、便捷及成本低的特点，已广泛应用于流行性细菌、病毒的定性定量检测等领域。本研究对 LAMP 的原理、特性及其应用做一综述，从而为其在食源性致病菌检测应用中提供理论依据。

Loop-mediated isothermal amplification (LAMP) technology is a new technology for nucleic acid amplification, which can amplify the specific target sequences to 109 level under the conditions of constant temperature at 60~65℃within 1h. It has the characteristics of high specificity, sensitivity, simplicity, conven-ience and low cost. LAMP has been widely used in qualitative and quantitative detection of bacterial and viral epidemic.This article reviewed the principle, characteristics and application of LAMP technology, in order to provide theoretical basis for its application in the detection of foodborne pathogens.

着重对近期关于新布尼亚病毒基本特性、国内外流行病学特点、检测方法等方面的研究进展进行综述。应用原子力显微镜观测病毒的基本形貌获得相关数据;流行病学方面SFTS病例也陆续在日本、韩国等国家有发现和报道;检测方面除针对病毒不同基因组的荧光定量方法外,还有反转录环介导的等温扩增技术,以及通过将反转录交叉启动扩增和垂直流偶联快速准确的检测该病毒。

This paper reviewed Severe Fever with Thrombocytopenia Syndrome Bunyavirus (SFTSV) pathogenic characteristics and epidemiological characteristics on domestic and foreign research, and the research progress of detecting methods. AFM observed the basic morphology of virus and related data;SFTS clinical cases were found and reported in countries such as Japan and South Korea;there is a reverse transcription loop mediated isothermal amplification (RT-LAMP), and the reverse transcription cross priming amplification (RT-CPA) and vertical flow (VF) for detection of SFTSV.

根据ISAV的基因保守序列,利用LAMP Designer软件设计了6条引物,采用新型的环介导等温扩增设备进行扩增和检测,优化了反应条件,分析了所建立方法的特异性和灵敏度,并与RT-PCR和实时荧光RT-PCR进行比较。研究表明,该方法最适反应温度为64℃,反应10 min就可以观察到明显的扩增。该方法灵敏度高,检测限为78.4 fg RNA,比常规RT-PCR灵敏度高100倍,与实时荧光定量RT-PCR灵敏度相当;特异性好,与传染性胰腺坏死病毒（IPNV）、鲤春病毒血症病毒（SVCV）、出血性败血症病毒（VHSV）、鱼类病毒性神经坏死病病毒（VNNV）、鱼腹水病毒（YAV）等14种主要鱼类病毒没有交叉反应。结果表明,本研究建立了ISAV的实时荧光环介导等温扩增检测方法,实验能对整个扩增过程进行实时监测,提高检测灵敏度的同时,防止由于开盖跑电泳或加染料而导致的污染。

The infectious salmon anaemia virus (ISAV) is classified as an Orthomyxoviridae. Its genome consists of 8 single-stranded negative-sense RNA segments. ISAV is the pathogen of fatal ISA listed by the World Organization for Animal Health (OIE). It mainly affects salmon farming in Europe and Northern America, but there has been a high chance of its introduction into China due to the increased salmon importation. Therefore it is very important to establish a rapid and accurate method for ISAV detection. Conventional ISAV detection methods involve cell isolation followed by RT-PCR or real-time RT-PCR. Recently Japanese scientists have established a novel technique with high sensitivity and rapidity, namely Loop-mediated isothermal amplification (LAMP) assay. In this study, LAMP assay was developed for detecting infectious salmon anaemia virus (ISAV). Six specific primers were designed according to ISAV genes using LAMP Designer software. A novel LAMP instrument was applied for the amplifica