环介导等温扩增联合横向流动试纸条检测简单异尖线虫/派氏异尖线虫方法的建立

简单异尖线虫复合种(Anisakissimplexspeciescomplex)是人兽共患寄生虫,其中的简单异尖线虫(A. simplex sensu stricto)和派氏异尖线虫(A. pegreffii)能引起人异尖线虫病。本研究利用环介导等温扩增技术(loop-mediatedisothermalamplification,LAMP)结合流动试纸条(lateralflowdipstick,LFD)建立了简单异尖线虫/派氏异尖线虫的快检技术。本研究以简单异尖线虫核糖体DNA的内转录间隔区(ribosomal internal transcribed spacer 2, rDNA-ITS2)序列为检测靶标,设计6条特异性的引物进行(荧光)LAMP反应。将生物素化的LAMP产物与异硫氰酸荧光素标记的探针杂交并通过LFD可视化显示。结果表明,本研究建立的LAMP-LFD可以特异性检测出简单异尖线虫/派氏异尖线虫,而对其它10种蠕虫的检测结果呈阴性;针对两个异尖线虫姊妹种的第三期幼虫(third-stagelarvae,L3)的检测灵敏度为单条虫体基因组DNA的10–5倍。优化后的LAMP反应时间为40min,加之探针杂交和LFD显色,整个检测时程只需50min。利用本LAMP-LFD方法对2015—2017年收集于140尾海鱼的1573条线虫进行检测的结果表明,简单异尖线虫/派氏异尖线虫是东海海域的优势种,这与经rDNA-ITS1-5.8S-ITS2测序的结果一致。因此, LAMP-LFD方法能快速、灵敏且准确地检测简单异尖线虫/派氏异尖线,在经济鱼类中简单异尖线虫复合种的筛检和常规检验检疫具有巨大潜力。     

基于LAMP-LFD技术的有害赤潮藻东海原甲藻(Prorocentrum donghaiense)快检方法

以东海原甲藻的ITS序列(Internal transcribed spacer,ITS)为检测靶标,将生物素标记的环介导恒温扩增技术(Loop-mediated isothermal amplification,LAMP)扩增产物与经异硫氰酸荧光素(Fluorescein isothiocyanate,FITC)标记的探针特异性杂交,并结合横向流动试纸条(Lateral flow dipstick,LFD)肉眼直接观察检测结果,建立了有害赤潮藻东海原甲藻(Prorocentrum donghaiense)的快速检测技术。经优化后的最适条件为63°C、30min,较常规PCR扩增缩短约2h。结果表明:LAMPLFD可特异性检出东海原甲藻,对常见赤潮藻检测结果均为阴性;其对东海原甲藻基因组DNA的检测最低限为47pg/μL,是常规PCR技术(以F3/B3为引物)的10倍。LAMP-LFD技术能高效、特异地检出东海原甲藻,仪器设备依赖性低,结果可视化,有望成为赤潮原因种检测监控的常规方法。     

考虑不同支架形式放大效应隔离开关地震响应分析

高压隔离开关作为变电站系统中的关键设备,提升其抗震性能对变电站的稳定性具有重要意义.通过Abaqus建立有限元模型,运用增量动力时程分析方法,研究了110kV隔离开关固有频率、放大系数随结构支架截面、高度和和瓷套管材料的变化规律.同时进一步研究了不同支架尺寸参数和不同瓷套管材料下隔离开关易损性特性.结果 表明:支架截面...     

梁铰型屈服RC框架结构地震弹塑性位移增大系数研究

本文通过弹性和弹塑性时程分析,研究了水平地震作用下梁铰型屈服RC框架模型结构的楼层屈服剪力系数、基本自振周期、楼层数3个因素对弹塑性位移增大系数的影响,通过非线性回归分析给出了弹塑性层间位移增大系数经验公式;通过分析滞回耗能沿楼层高度的分布,初步确定了梁铰型屈服RC框架结构的薄弱楼层位置;基于结构损伤分析,讨论了抗震规范中RC框架结构弹塑性层间位移角限值的水准。     

设置局部粘滞消能支撑的框架节点抗震试验研究

提出了具有位移放大效应的局部粘滞消能支撑的构成及工作原理,开展了设置该类支撑的钢筋混凝土梁-柱节点的反复加载试验,初步考察了节点的抗震性能,并通过对比试验与常规钢筋混凝土梁-柱节点进行了比较。研究结果表明,安装该类支撑后节点的耗能能力明显提高,梁端位移幅值较大时提高幅度可达200%以上,大大改善了节点的抗震性能。     

Experimental study of the ground motion on a large scale topographic hill at Kitherion (Greece)

We present the results of an experiment about the effects on the seismic ground motion of a hill of large dimensions (6km long, 3km large and 700m high) near Corinth (Greece). We installed 7 seismometers across the hill, and analyzed the ground motion with different methods: the classical spectral ratios (CSR) and the horizontal to vertical spectral ratios calculated both on noise (HVNR) and earthquake data (RF). The amplification at the resonant frequency (around 0.7Hz) is low, as expected, but one of the two stations installed at the top exhibits large amplification (up to a factor of 10) at 3Hz. The comparison between the H/V results and the CSR ones suggests that the former method is able to point out the fundamental frequencies of a hill. Amplifications and spatial localization predicted by numerical 3D modeling are consistent with the experimental data only at frequencies lower than 1Hz.     

Applications of the Mild-Slope Equation to Tidal Computations in the Taiwan Strait

Employing harmonic analysis of tidal data in the Taiwan Strait, the cross-strait tidal characteristics are completely illustrated. Based on the two dimensional mild-slope equation which can be reduced to the shallow-water wave equation, a finite element model (Tsay et al., 1989) is applied to investigate the characteristics of tides in the Taiwan Strait. The co-range and equi-phase charts of major tidal constituents, such as M₂, S₂, N₂, and K₁, are reproduced. Anomalous amplification of semidiurnal tides in the Taiwan Strait is verified. With rotation effects neglected and by applying a non-reflective condition on the open boundaries, the numerical results of phase-lag and co-range distributions show very good agreement with observed data for semidiurnal tides in the Taiwan Strait. Due to crude representation of the topography at two ends along the China coast, computed tidal distributions deviate from the observations. However, both computed amplitudes and phase-lags compare very well with observed data along the central half of the China coast.     

EST-derived microsatellites in Pseudosciaena crocea and their applicability to related species

Simple sequence repeat (SSR) markers were obtained for the large yellow croaker Pseudosciaena crocea using 1 205 expressed sequences tags (ESTs) from the NCBI database. Primers for 48 ESTSSR loci were designed and screened with 30 P.crocea specimens captured from Guanjingyang sea area in Fujian Province of China. Sixteen of the loci were polymorphic, which were amplified with 3 to 11 alleles per locus and the mean of 6.13. The observed and expected heterozygosity per locus ranged from 0.091 to 0.844 (mean 0.544) and from 0.118 to 0.892 (mean 0.644), respectively. Polymorphic information content (PIC) ranged from 0.115 to 0.866 (mean 0.593). The results for cross-species amplification of the 16 large yellow croaker EST-SSRs on P. polyactis, C. niveatus, C. lucidus, A. argentatus and J. belengeri revealed that 14, 12, 11, 7 and 6 loci were successfully amplified with 1 to 10 alleles with an average of 4.5 per locus, respectively, which are suitable for population genetics studies of these species and useful for phylogenetic relationship analysis among these species. Overall, this study provides a set of type I markers for population genetics studies and genome mapping for large yellow croaker and its closely related species.     

环介导恒温扩增技术快速检测米氏凯伦藻方法的建立

米氏凯伦藻(Karenia mikimotoi,Hansen)属于裸甲藻目(Gymnodiniales),裸甲藻科(Gymnodi-niaceae),凯伦藻属(Karenia)。它能产生溶血性(hemolytic)毒素和鱼毒(ichthyotoxins),溶解鱼类细胞,破坏鱼鳃组织结构,使鱼类无法正常呼吸而窒息死亡[1]。近年来,米氏凯伦藻引起的赤潮在世界各海域屡次发生,日本海域,墨西哥湾,新西兰、韩国、苏格兰和澳大利亚海域都有米氏凯伦藻引发赤潮的报道,对渔业造成很大损失[2]。