A rapid loop-mediated isothermal amplification method for detection of the modified GM cry1A gene in transgenic insect-resistant cotton and rice

Among the commercial genetically modified (GM) crops, the insect-resistant GM crops are the major cultivars that cry gene is introduced into. A cry1Ab/1Ac GM fusion gene (GFM cry1A) and a GM truncated cry1Ac gene (cry1Ac-Mon) is the key foreign gene employed for construction of GM crops by China researchers and Monsanto Technology LLC respectively. Here these two genes are entitled “GM cry1A” gene and a rapid visual loop-mediated isothermal amplification (LAMP) assay method for detection of GM cry1A in transgenic insect-resistant crops was established. The LAMP assay was performed at an optimal temperature of 65 °C for 60 min in the presence of a set of four specific primers recognized six distinct sequences of the GM cry1A gene. The rough detection limit to the GM cry1A in samples is as low as 0.01% (a weight ratio of transgenic insect-resistant rice/cotton to non-transgenic rice/cotton). Comparatively, the sensitivity of this LAMP method is 10 times over that of the conventional PCR method. Fifteen cultivars/events and five Bt strains with or without cry1A gene were analyzed using the LAMP method as well as PCR method. The results demonstrate that this LAMP method shows a distinct specificity to the GM cry1A gene comparing with PCR analysis. Therefore, this LAMP method will be a potential effective tool for screening the GM cry1A gene in GM crops which are widely plant in China and other developing countries.     

A novel developed method based on single primer isothermal amplification for rapid detection of Alicyclobacillus acidoterrestris in apple juice

Alicyclobacillus acidoterrestris (A. acidoterrestris), as one of the most major spoilage-causing species within the Alicyclobacillus genus, can survive and multiply in the pasteurization process. Nowadays, A. acidoterrestris has become worldwide issue in the fruit juice industry due to its spore-forming and thermo-acidophilic features. A novel single primer isothermal amplification assay (SPIA) was developed for the rapid detection of A. acidoterrestris in apple juice. This assay was designed to target the 16S rRNA gene with a DNA/RNA chimeric primer. Detection of gene amplification was accomplished by amplification curve, turbidity and addition of single strand DNA binding dye SYBR Green II allowing visualization under ultra violet light. The specificity of the assay was confirmed using 7 strains of A. acidoterrestris and 30 strains of non-A. acidoterrestris. The SPIA was highly sensitive and the detection limit was as low as 4.8 CFU/mL. Furthermore, the apple juice inoculated with 61 CFU/mL of A. acidoterrestris could be detected as positive. The novel SPIA with visualization results was successfully applied for the detection of A. acidoterrestris and exhibited high specificity and sensitivity, making it a powerful tool for the detection of A. acidoterrestris in fruit juice industry and being conveniently applied in developing countries with limited resource.     

多层石膏墙体钢网格式框架结构影响系数研究

GB 50011-2010《建筑抗震设计规范》并未给出多层石膏墙体钢网格式框架结构的结构影响系数,因此其抗震设计荷载取值还不明确。基于此,依据GB 50011-2010规范设计了6个典型的多层钢网格式框架结构模型,通过推覆分析得出结构的能力谱曲线,基于改进能力谱法得出罕遇地震和抗震设防烈度地震作用下的需求谱曲线。将结构的能力谱曲线和延性需求谱曲线族置于统一坐标系中,确定结构的目标位移以及性能关键点坐标。进一步考虑层数及标准层布置对结构延性和超强系数的影响,评估结构的抗震性能并给出结构影响系数取值。研究表明：多层钢网格式框架结构在抗震设防烈度地震和罕遇地震作用下的变形均满足抗震规范要求;其结构影响系数取3.0,位移放大系数取4.0;该结构可采用现行抗震规范进行设计,但设计结果偏于保守。     

顺层岩质边坡加速度响应规律和滑动堵江机制大型振动台试验研究

根据动力模型试验的相似关系,制作1个尺寸为3.0 m×0.8 m×1.6 m (长×宽×高)坡角为40°的顺层模型边坡,设计传感器的布置和试验输入方案,并完成大型振动台试验。试验发现,在坡面上水平加速度随着坡高增加有明显的放大作用,并且该放大作用呈现出非线性和在不同高程的分段性特征,在坡高的1/3以下,加速度放大效应明显,在坡高2/3到坡顶处放大系数急剧增大,中间段水平加速度放大系数增长较为缓慢;在坡体内部竖直方向上,随着高程增加,加速度放大系也表现出非线性放大效应;在坡体水平方向上,在坡面一定深度内坡面处的加速度放大系数大于坡体内部,表现出趋表效应。根据破坏后的特征及试验过程中的视频回放和分析,得出模型边坡的滑动堵江机制为地震诱发→后缘松动拉裂→前缘剪切→高速下滑→对岸阻挡隆起→堆积坡脚→形成堰塞体。研究成果对地震灾区滑坡以及堰塞湖形成机制的认识和减灾防灾有一定的价值。     

基于柔性铰链的柔性放大机构参数化设计

为了对柔性微位移放大机构进行优化设计,有必要对柔性铰链及柔性放大机构进行参数化分析与研究。提出了一个通用的结构参数ε,探讨了ε对不同柔性铰链柔度系数的影响规律,并横向比较了常用柔性铰链的柔度特性。另一方面,基于柔度特性的影响分析,提出了新的参数柔度比λ,重点分析了不同柔度比λ的柔性铰链主要输出位移形式的灵敏度。以实际的桥式柔性微位移放大机构为例,利用参数ε和λ实现了该柔性放大机构的参数化设计,并用有限元软件进行了仿真计算。实验测量结果表明,对基于柔性铰链的柔性微位移放大机构进行参数化设计,最终输出位移行程与有限元仿真设计的结果误差率为3.80%。基于柔性铰链的结构参数ε和柔度比λ对柔性放大机构进行参数化设计是可行且正确的,有利于这一类柔性放大机构的优化设计。     

A Novel Isothermal Assay of Borrelia burgdorferi by Recombinase Polymerase Amplification with Lateral Flow Detection

A novel isothermal detection for recombinase polymerase amplification with lateral flow (LF-RPA) was established for Borrelia burgdorferi (B. burgdorferi) detection in this study. This assay with high sensitivity and specificity can get a visible result without any additional equipment in 30 min. We designed a pair of primers according to recA gene of B. burgdorferi strains and a methodology evaluation was performed. The results showed that the RPA assay based on the recA gene was successfully applied in B. burgdorferi detection, and its specific amplification was only achieved from the genomic DNA of B. burgdorferi. The detection limit of the new assay was about 25 copies of the B. burgdorferi genomic DNA. Twenty Lyme borreliosis patients’ serum samples were detected by LF-RPA assay, real-time qPCR and nested-PCR. Results showed the LF-RPA assay is more effective than nested-PCR for its shorter reaction time and considerably higher detection rate. This method is of great value in clinical rapid detection for Lyme borreliosis. Using the RPA assay might be a megatrend for DNA detection in clinics and endemic regions.     

适配体功能化的DNA水凝胶的制备及其在食品安全小分子快速检测中的应用

DNA水凝胶是具有三维聚合物网络的高保水性材料。研究人员设计了多种DNA水凝胶交联制备方法,并通过向其中引入其他功能分子或与其他类型的功能材料相互结合,构建了具有优异性能的DNA水凝胶,受到了广泛关注。适配体是基于指数富集的配体系统进化(systematicevolutionofligandsbyexponential enrichment,SELEX)技术从随机寡核苷酸文库中筛选获得的对目标物质具有良好特异性与亲和力的寡核苷酸序列。适配体功能化的DNA水凝胶具有靶向范围广、稳定性好、易于修饰、操作简单和成本低等优点,得到了广泛应用。本文概述了构建适配体功能化的DNA水凝胶的基本设计原则与分类,重点介绍了适配体功能化的DNA水凝胶在食品安全检测中的最新策略,最后,讨论了适配体功能化的DNA水凝胶面临的挑战以及对未来的展望,旨在为其在食品安全领域的应用提供参考。     

超短中红外激光脉冲的产生及其发展状况

超短中红外脉冲对研究物理、化学、生物学和半导体内的超快过程等有非常重要的作用。介绍了参量振荡、差频产生、参量放大产生超短中红外激光脉冲的方法,及国内外相关研究的最新进展。