排序方式: 共有63条查询结果,搜索用时 31 毫秒
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目的建立可靠的临床血清(浆)循环miRNAs定量技术。方法常规收集血清(浆)标本,mirVana PARIS试剂盒法抽提血清(浆)总RNA,采用DNaseI消化总RNA提取液,以miRNAs特异性茎-环引物引导反转录,通过TaqMan实时荧光定量PCR对U6及靶miRNAs进行检测。结果10份新鲜血浆总RNA浓度介于3.5~35.4ng/μl之间。对常规收集的400μl临床血清标本中U6、miR-16、miR-224均能实现特异扩增及定量,相应的平均Ct值约为30、25及32。6份不同留置时间血清标本总RNA浓度分别10.24和4.46ng/μl,定量PCR结果显示其中相应miR-16和miR-224的丰度却相对稳定。结论血清(浆)总RNA抽提及循环miRNAs定量切实可行。 相似文献
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Juan García-Bernalt Diego Pedro Fernndez-Soto Antonio Muro 《International journal of molecular sciences》2022,23(22)
Since the onset of the COVID-19 pandemic, over 610 million cases have been diagnosed and it has caused over 6.5 million deaths worldwide. The crisis has forced the scientific community to develop tools for disease control and management at a pace never seen before. The control of the pandemic heavily relies in the use of fast and accurate diagnostics, that allow testing at a large scale. The gold standard diagnosis of viral infections is the RT-qPCR. Although it provides consistent and reliable results, it is hampered by its limited throughput and technical requirements. Here, we discuss the main approaches to rapid and point-of-care diagnostics based on RT-qPCR and isothermal amplification diagnostics. We describe the main COVID-19 molecular diagnostic tests approved for self-testing at home or for point-of-care testing and compare the available options. We define the influence of specimen selection and processing, the clinical validation, result readout improvement strategies, the combination with CRISPR-based detection and the diagnostic challenge posed by SARS-CoV-2 variants for different isothermal amplification techniques, with a particular focus on LAMP and recombinase polymerase amplification (RPA). Finally, we try to shed light on the effect the improvement in molecular diagnostics during the COVID-19 pandemic could have in the future of other infectious diseases. 相似文献
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《International Journal of Hydrogen Energy》2022,47(76):32455-32472
Sugarcane bagasse (SCB) pre-treated with laccase was used as substrate in two acidogenic phases (1 P and 2 P). In 1 P, the pretreatment conditions were statistically optimized (pH 4.0, 37 °C, 0.4 U of laccase/mL, 24.0 g SCB/L and 120 min of reaction), which allowed obtaining 84% more H2 (166.8 mL/L) than SCB in natura (90.4 mL/L). In 2 P, the liquid fraction of the 1 P reactors was diluted (90%) and more 108.5 ml H2/L was obtained. Furthermore, the quantification of enzymes genes responsible for the hydrolysis (Cel) and H2 production (Hyd) was carried out. An increase in the expression of both genes was observed from the phase of highest H2 production rates in P1. In 2 P, the 1 P low genic expression was indicative of the autochthonous bacteria activity. The 2nd acidogenic phase strategy proved to be a promising novelty for the use of pre-hydrolyzed substrates with concomitant production of more value-added products. 相似文献
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实时荧光定量聚合酶链式反应是新型冠状病毒COVID-19快速检测的重要手段,病毒样本经过一系列复杂的化学反应处理后通过RT-qPCR温控系统得到检测结果,RT-qPCR温控仪实际就是一台精准的测温、控温仪和光谱检测设备。本文采用ARM处理器+H桥驱动器+帕尔贴的设计架构,通过归一化PID算法解决温度的快速升降问题,从而构建低成本、小型化的温控系统。该系统可以根据不同的温度设定和荧光剂选择适用其它病毒的核酸检测,整体控温指标可以达到升温速度不小于15℃/s、降温速度不小于8℃/s、温度精度0.1℃以内、升降温过冲小于4℃,稳定时间3s。 相似文献