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以3种海鱼(牙鲆、狭鳕、鲱鱼)为样本,针对海产品中主要的致病性寄生虫-异尖线虫,开展了酶消化检测技术的研究.根据鱼肉消化前后干物质的质量变化设计了胃蛋白酶消化效率的计算方法,并按照鱼肉:消化液＝1:10 (g/mL)的反应比例,分别确定了酶水解鱼肉的最佳条件为:初始pH值为1.1,温度为37 ℃,酶活力为8 U/mL左右;消化后所得的虫体采用多重PCR方法替代传统的形态学观察进行种属鉴定,从而初步建立了基于酶水解-多重PCR的异尖线虫的酶消化检测体系.实际样本检测表明,该技术可以较为准确、快速地用于海产鱼类中简单异尖线虫(Anisakis simplex)和伪地新线虫(Pseudoterranova decipiens)的确证性检测. 相似文献
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自动气象站观测系统在气象领域中应用广泛,数据采集器是系统中的关键部件,改进了低功耗、多功能、多通道、智能化控制等性能.传感器接口包括DAC实现的可调恒压源和可调恒流源、稳压二极管实现的前端保护、多路模拟开关实现的通道控制和测量类型控制、精密线性光耦合器实现的隔离电路、DAC和仪表放大器实现的增益控制等硬件部件.设计单片机程序实现电路的各种控制及要求功能.经制板、调试,该数据采集器能正常运行. 相似文献
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Although Escherichia coli is widely distributed in the marine environment, only a small percentage are pathogenic to humans. Nonetheless, the widespread occurrence of waterborne infections of E. coli origin in humans has become one of the major health problems worldwide. To date, several types of enterovirulent E. coli have been recognized as the aetiologic agents of various gastrointestinal infections in humans. The most commonly encountered are those belonging to the enterotoxigenic (ETEC), enteroinvasive (EIEC), enterohaemorrhagic (EHEC) and enteropathogenic (EPEC) subtypes. In order to better determine the health risks that are associated with exposure to some of these specific subtypes, we have developed a very sensitive multiplex PCR system for the rapid detection and typing of ETEC, EHEC and possibly EPEC strains of E. coli in the aquatic environment. The target genes chosen for this investigation included: the PHO-A housekeeping gene (present in all E. coli); the LT1, LT2 and ST1 genes of ETEC; the VT1 and VT2 verotoxin, and EAE virulence genes of EHEC and EPEC, respectively. Six pairs of oligonucleotide primers were designed to simultaneously amplify internal fragments of these genes by multiplex PCR to generate PCR products that could be analysed and confirmed with relative ease by gel electrophoresis and HincII enzyme digestion. The results showed that the six sets of PCR primers were highly specific for their target genes and produced specific amplimers of the expected size from several control strains of E. coli – ATCC 35401 (LT1+/ST1+); SA53 (LT2+/VT2+); and O157 (VT1+/VT2+/EAE+). The detection sensitivity of the multiplex PCR system for the six target genes in an E. coli cell mixture was optimized and enhanced by preincubating serially diluted cells in Luria-Bertani broth for 6 h prior to PCR analysis. The results obtained indicated a detection sensitivity of 10° CFU (of each strain) per 100 μl reaction. Multiplex PCR analysis of seawater samples collected from four sewage-polluted sites in Hong Kong indicated the presence in all four samples of E. coli bacteria that were positive for LT1, ST1, VT1 and EAE virulence genes. Overall, the data indicated that the multiplex PCR system described in this study is a potentially very useful and powerful method for routine monitoring and risk assessment of water quality. 相似文献
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用8个微卫星标记组合建立了2个微卫星多重PCR体系,对大菱鲆7个人工选育家系进行了系谱认证、亲子鉴定和遗传多样性研究。2个多重PCR体系中8个微卫星位点共检测到等位基因49个,每个位点等位基因数为3~8个。根据已知亲本及子代基因型,成功推断出了3个缺失亲本的基因型。在双亲未知的情况下2个多重PCR的8个微卫星位点累积排除概率是96.58%,已知1个亲本时累积排除率为99.71%,亲子鉴定准确率为96.42%。采用70个个体进行双盲验证,利用UPGMA法对7个家系的70个体进行了聚类分析,同一家系95.71%的个体聚类分析结果与系谱来源一致。Cervus 2.0软件亲子鉴定结果表明亲子鉴定准确率为92.86%。2个多重PCR体系的建立为大菱鲆不同家系混养后的亲子鉴定、系谱分析和分子辅助家系管理提供了技术手段。 相似文献
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二温式多重PCR检测鉴别对虾白斑综合征病毒(WSSV)和传染性皮下及造血器官坏死病毒(IHHNV)的研究与应用 总被引：7，自引：0，他引：7
建立了一种同时检测对虾白斑综合征病毒（WSSV）、传染性皮下和造血器官坏死病毒（IHHNV）的二温式多重PCR技术。根据基因库中WSSV（AF369029）和IHHNV（NC002190）基因序列，设计了两对分别与WSSV和IHHNV某段保守基因序列互补的引物，用这两对引物对同一样品中的WSSV和IHHNVDNA模板进行多重PCR扩增，结果均同时得到了两条大小与实验设计相符的593bp（WSSV）和356bp（1HHNV）特异性多重PCR扩增条带，而对其他对虾疾病病原的PCR扩增结果均为阴性；敏感性测定结果表明，该多重PCR技术最低能检测到WSSV和IHHNVDNA各100pg。用该多重PCR对400份临床病料进行检测，结果230份检出WSSV，阳性率57．5％；96份检出IHHNV，阳性率24％；56份同时检出WSSV和IHHNV，阳性率14％。18份为阴性，阴性率为4．5％。结果提示了WSSV和IHHNV广泛存在于中国南方的养殖对虾中，作者建立的二温式多重PCR可以用于这两种病毒的临床快速检测和鉴别诊断。 相似文献
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筛选多态性高、特异性好、片段大小适中的10 个长牡蛎微卫星位点进行优化组合, 根据扩 增片段大小及同一荧光不重叠原则构建了两组五重PCR 体系。运用CERVUS 3.0 软件对0527 组27 个长牡蛎全同胞家系的643 个子代和0612 组27 个全同胞家系382 个子代分别进行亲权鉴定。结果 发现, 用两组微卫星五重PCR 鉴别时, 在0527 组和0612 组的鉴定成功率均为100%; 只用第一组微 卫星五重PCR, 可以将0527 组96%的子代和0612 组96%的子代鉴定到亲本; 只用第二组微卫星五 重PCR, 可以将0527 组97%的子代和0612 组95%的子代鉴定到亲本。本研究中筛选出的两组微卫 星五重PCR 体系在两组家系中的鉴定效率均较高, 可以快速有效地将子代个体鉴定至所属父母本, 在长牡蛎家系鉴定中具有很好的应用价值。 相似文献
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Quy Thy Truong Cyril de Runz Guillaume Touya 《International journal of geographical information science》2019,33(8):1651-1682
This paper aims to qualify the behaviour of contributors to OpenStreetMap (OSM), a volunteered geographic information (VGI) project, through a multigraph approach. The main purpose is to reproduce contributor’s interactions in a more comprehensive way. First, we define a multigraph that combines existing spatial collaboration networks from the literature with new graphs that illustrate collaboration based on specific aspects of the VGI modes of contribution through semantics, geometry and topology. Indeed, the ways that contributors interact with one another through editing, completion, or even consumption may provide additional information on each user’s operation mode and therefore, on the quality of the contributed data. Social collaborations drawn from indirect criteria – for example, comparisons between contributors’ activity areas – can also be contemplated under another network. Second, the resulting multigraph is analysed using data mining approaches to characterise individuals and identify behavioural groups. The implementation of a multiplex network based on an OSM data sample and an initial analysis make it possible to identify useful behaviours for data qualification. The initial results characterise some contributors as pioneers, moderators and truthful contributors, according to their special roles in the graphs. Mapping elements that include these contributors’ participation are likely to be reliable data 相似文献
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福建沿海巨蛎属牡蛎的主要种类及其分布 总被引：4，自引：0，他引：4
本实验采用了多重种类特异性PCR（multiplex species—specific PCR）技术,研究了巨蛎属（Crassostrea）牡蛎主要种类在福建沿海的分布．从沿海11个采样地点共采集了657个野生牡蛎样本,随机抽取了327个牡蛎样本进行基因组DNA的提取和线粒体COI基因的鉴定,结果发现200个个体为葡萄牙牡蛎（Crassostrea angulata）,101个个体为近江牡蛎（Crassostrea ariakensis）,20个个体为熊本牡蛎（Crassostrea sikamea）,6个个体为香港巨牡蛎（Crassostrea hongkongensis）．此次实验中未发现长牡蛎（Crassostrea gigas）．结果表明,福建沿海有葡萄牙牡蛎、近江牡蛎、熊本牡蛎和香港巨牡蛎4种巨蛎属牡蛎分布． 相似文献
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洪积扇相砂砾岩体储层构型研究方法探讨 总被引：6，自引：1，他引：5
在厘定其不同层次界面成因的基础上,把Miall研究思路成功移植到洪积扇相复杂储层中,建立了与开发精度相匹配的洪积扇储层内部结构研究的层次分析法,即复合水道、单期水道和高渗透段3个层次。划分出洪积扇砂砾岩储层5级构型界面:5级界面为洪泛沉积发育稳定的泥岩,构成油田开发中的隔层;4级界面由厚度变化的泥岩、粉砂质泥岩组成,形成流体渗流屏障;3级界面为形成的支撑砾岩体与周围储集体的岩性突变面。总结了复合水道规模扇缘亚相单期条带状、扇中亚相多期交织的宽带状及扇顶亚相多期垂向叠加的厚层板状砂砾岩体的分布特点。提出了洪积扇单期水道研究方法,即分别从单期水道4个方面的垂向识别特征和4种单期水道的侧向界面识别标志入手,在3种单期水道空间组合模式指导下开展三维空间组合研究。2个典型井区的实例分析表明,扇顶亚相单期水道的边界沿着古水流方向近似呈直线分布,多期水道呈楔形叠瓦状前积叠加,垂直物源方向为垂向加积切割叠置关系;扇中亚相单期水道边界类似曲率较小的辫状河道边界,多期水道具有侧向叠置的特点。 相似文献