共查询到20条相似文献,搜索用时 62 毫秒
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近年来,越来越多新的致病性呼吸道病毒被发现,给口岸检验检疫机构的预防和控制造了成极大困扰,建立甲型流感病毒(Influenza A virus,FluA)、乙型流感病毒(Influenza B virus,FluB)、副流感病毒(Human Parainfluenza virus,HPIV)、冠状病毒(Coronavirus,Cov)、呼吸道合胞病毒(Respiratory syncytial virus,RSV)、人偏肺病毒(Humanmetapneumovirus,HMPV)、人博卡病毒(Human bocavirus,Hbov)、腺病毒(Adenovirus,Adv)和鼻病毒(Rhinovirus,Rhv)9种呼吸道病原体微流体芯片检测体系。选择人β-actin基因作为靶基因设计内标引物探针,根据9种呼吸道病原体基因组的保守区设计相应的引物探针;在Ultra Fast Labchip Real-time PCR V280平台上,筛选合适的RT-PCR Master Mix;对扩增体系中引物探针浓度进行优化,用优化后的扩增体系进行灵敏度和特异性验证;用100例临床标本进行回顾性检测... 相似文献
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基于MEMS技术的毛细管电泳芯片 总被引:1,自引:0,他引:1
基于MEMS技术的毛细管电泳芯片具有体积小、成本低、便于携带、分析速度快、所需样品和试剂少等优点,是一种新型的微型分析实验装置。介绍该芯片的结构、进样方式、材料选取、制作工艺、键合方法、样品检测方法等方面的研究进展和存在的问题,展望了应用前景 。 相似文献
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八种动物血清蛋白的聚丙烯酰胺凝胶电泳和在进化中相互的关系 总被引:3,自引:0,他引:3
本文利用聚丙酰胺凝胶电泳方法分析了隶属两个纲6个目的8种动物血清蛋白。它们是两栖纲的青蛙、大鲵;哺乳纲的树鼩、长爪沙鼠、穿山甲、豹猫、狢和果子狸。 在分析了科以下分类阶元各种动物血清蛋白组成及进化的基础上,我们比较了这8种亲缘关系较远并在分类上隶属于各不同高级分类阶元之间的关系。 分析结果表明,在纲与纲、目与目之间,动物血清蛋白中特有蛋白的比例数,随起源时间由远到近,有增大的趋势;而在科与科之间则相反。共有蛋白区带的比例随着分析范围的扩大而逐渐变小。 这些结果为纲、目及科的分类学的宏观定义提供了分子水平上可以参考的实验依据。 相似文献
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本文叙述一个新设计的微电极放大器,能用同一根微电极同时进行微电泳(如注射 HRP对神经元进行染色)和记录生物电。其主要性能如下:(1)输出到微电极的电压的范围为±50V。(2)在整个电压范围内能对杂散输入电容进行补偿。(3)微电泳的电流恒定,不受微电极电阻变化的影响。(4)注射电流时放大器工作正常,具有极高输入阻抗,低栅流和低噪声等特性。 相似文献
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一种简单快速的聚丙烯酰胺凝胶电泳染色法——铜染色法 总被引:2,自引:0,他引:2
十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDSPAGE)作为分离鉴定生物大分子的有效手段已广泛用于分子生物学和医学临床工作,迄今为止,用于该技术的染色法多沿用考马斯亮蓝(CBB)和银染色。但上述两法均有其局限性,如蛋白凝胶染色前须经酸醛固定,不易洗脱且操作繁琐,脱色耗时。最近,Lee等报道一种SDS-PAGE负染色法,利用氯化铜分别与凝胶中 相似文献
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六种疫霉菌体可溶性蛋白质和酯酶的聚丙烯酰胺凝胶平板电泳研究 总被引:14,自引:0,他引:14
本文对槐生疫霉、掘氏疫霉、樟疫霉、烟草疫霉、棕榈疫霉及辣椒疫霉等6种疫霉81株菌菌体可溶性蛋白质和酯酶进行了聚丙烯酰胺凝胶平板电泳研究。结果指出,种间蛋白质图谱差异明显,种内菌株间基本一致。就酯酶图谱来说,种内菌株间存在一定差异,但种间差异更为显著。因此,本试验进一步支持槐生疫霉新种的建立。同时表明,菌体可溶性蛋白质电泳图谱在疫霉种的鉴定和分类上具有重要意义,而酯酶电泳图谱在一定程度上也有助于疫霉 相似文献
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低温保存对卵母细胞造成渗透损伤、毒性损伤和冰晶损伤,使得细胞冻后质量难以提高.本文首次提出将微流控法添加-去除保护剂分别与三种冷冻载体(OPS、QC及Cryotop)搭配使用,对猪卵母细胞进行冷冻保存,并与传统冷冻法进行比较;然后,首次选用透明陶瓷和玻璃制作集成一体化芯片,对猪卵母细胞进行冷冻保存,以冷冻保存后的细胞存活率和发育率为判断依据,筛选出较好的方案;最后,对冻后卵母细胞的早期凋亡情况、胞内活性氧水平和线粒体膜电位水平进行分析.结果表明,微流控添加-去除保护剂组卵母细胞冻后存活率以及卵裂率都显著高于传统冷冻组,可以有效降低卵母细胞的早期凋亡率和胞内活性氧水平,减小线粒体损伤,提高细胞的冻后质量.透明陶瓷一体化芯片保存卵母细胞得到的存活率和卵裂率与传统OPS冷冻的保存结果无显著差异.微流控芯片技术为卵母细胞的低温保存提供新的思路,有较好的应用前景. 相似文献
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目的 构建Y染色体短串联重复序列(Y-STR)微流控芯片扩增检测试剂,并进行性能验证,实现Y-STR基因座的快速全集成检测。方法 使用Y-STR微流控芯片检测体系,对其灵敏度、成功率和分型准确率、峰平衡性、精准性和准确性、检材适应性、混合物检测能力和抗抑制性进行验证评估。结果 DNA标准品9948的模板量≥8 ng,血卡片数≥3片以及口腔拭子刮擦次数≥7次时可获得Y-STR完整分型;165份样本的全集成检测成功率为91.52%,分型准确率为99.74%;不同荧光通道之间的峰高比值为89.81%;10次运行的等位基因分型标准品的片段大小标准差均在0.5 bp以内,20份样本全集成检测的等位基因片段和相应的等位基因标准品之间的片段准确性均在0.5 bp以内;能够对口腔拭子、血卡、唾液卡、烟蒂、血棉签、布片精斑等检材进行准确分型;混合样本中较小贡献者与较大贡献者在1∶3的比例时可获得完整基因分型;在不同浓度的腐殖酸(50~400 mg/L)、靛蓝(20~100 nmol/L)、血红蛋白(100~500μmol/L)等抑制物的干扰下,该体系可获得完整基因分型。结论 该体系可应用于国产Quick... 相似文献
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微流控芯片技术是一种全新的微量分析技术。介绍了微流控芯片技术的基本原理、特点及分类,并深入讨论了该技术在食品安全、营养、加工和风味等食品领域中的应用,包括有害化学物质、食品添加剂、转基因食品和食源性致病微生物等的检测,营养物质和功能成分的分析鉴定,食品工艺参数的调控以及食品风味成分的检测,展望了微流控芯片技术在食品领域的广阔应用前景。 相似文献
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描述了一种微流控芯片电泳快速分离血清高密度脂蛋白(high density lipoprotein,HDL)亚类的方法.利用自制的微流控芯片,结合激光诱导荧光检测系统,40mmol/L Tricine、50mmol/L甲基葡胺(MEG)、0.2mmol/LSDS(pH8.5)为样品缓冲液,40mmol/L Tricine、50mmol/LMEG、0.01mmol/L SDS(pH8.5)为分离缓冲液,4min内HDL3和HDL2两种亚类得到基线分离.该法操作过程简单,重复性较佳,测试费用低廉,在临床HDL亚类的检测中具有较好的应用前景. 相似文献
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目的:利用新型纳米森林材料,构建一种操作简单、检测快速、灵敏度高的用于现场检测的自驱动微流控芯片。方法:利用MEMS加工技术制备出具有优良光学性能和大表面积的石英纳米森林结构微流道,对该纳米森林结构的高度、宽度/横向尺寸、密度、表面积、光学性能、毛细驱动效果、荧光增敏效果做出评价,利用双抗体夹心的方法进行蓖麻毒素的检测。结果:纳米纤维锥底直径200~300nm,高度约1. 0μm,纳米森林的密度约为10个/μm~2,估测表面积比底面积达5∶1以上。其在波长为680nm处的透光率达89. 5%,驱动流速约5mm/s,与平面结构相比,其饱和荧光显色成倍提高。蓖麻毒素的检测限低于10pg/ml,在10~6 250pg/ml范围内具有较好线性关系。结论:基于纳米森林结构,成功构建了一种具有超大表面积和高灵敏度的毛细自驱动微流控芯片。 相似文献
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Liquid biopsies serve as both powerful noninvasive diagnostic tools for early cancer screening and prognostic tools for monitoring cancer progression and treatment efficacy. Exosomes are promising biomarkers for liquid biopsies, since these nano‐sized extracellular vesicles (EVs) enrich proteins, lipids, mRNAs, and miRNAs from cells of origin, including cancer cells. Although exosomes are abundantly present in various bodily fluids, conventional exosome isolation and detection methods that rely on benchtop equipment are time‐consuming, expensive, and involve complicated non‐portable procedures. As an alternative, recently developed microfluidic platforms can perform effective exosome separation and detection for liquid biopsies using a single device. Such methods offer advantages of integrity, speed, cost‐efficiency, and portability over conventional benchtop and early microfluidic‐based single‐functional methods which can only separate or detect exosomes separately. These advances have made exosome‐based point‐of‐care (POC) applications possible. This review outlines recent integrated microfluidic‐based exosomal detection strategies to guide future development of such devices for use in liquid biopsies for early cancer screening, prognostic monitoring, and other potential POC applications. 相似文献
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Klaus Eyer Phillip Kuhn Simone Stratz Petra S Dittrich 《Journal of visualized experiments : JoVE》2013,(80)
We present a microfluidic device that enables the quantitative determination of intracellular biomolecules in multiple single cells in parallel. For this purpose, the cells are passively trapped in the middle of a microchamber. Upon activation of the control layer, the cell is isolated from the surrounding volume in a small chamber. The surrounding volume can then be exchanged without affecting the isolated cell. However, upon short opening and closing of the chamber, the solution in the chamber can be replaced within a few hundred milliseconds. Due to the reversibility of the chambers, the cells can be exposed to different solutions sequentially in a highly controllable fashion, e.g. for incubation, washing, and finally, cell lysis. The tightly sealed microchambers enable the retention of the lysate, minimize and control the dilution after cell lysis. Since lysis and analysis occur at the same location, high sensitivity is retained because no further dilution or loss of the analytes occurs during transport. The microchamber design therefore enables the reliable and reproducible analysis of very small copy numbers of intracellular molecules (attomoles, zeptomoles) released from individual cells. Furthermore, many microchambers can be arranged in an array format, allowing the analysis of many cells at once, given that suitable optical instruments are used for monitoring. We have already used the platform for proof-of-concept studies to analyze intracellular proteins, enzymes, cofactors and second messengers in either relative or absolute quantifiable manner. 相似文献
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Comparative Proteome Analysis of Human Temporal Cortex Lobes by Two-Dimensional Electrophoresis and Identification of Selected Common Proteins 总被引:2,自引:0,他引:2
Human brain proteins were isolated from left and right temporal cortex lobes at the age of 73, 23, 84 years and separated by two-dimensional gel electrophoresis (2-DE). 2-DE was carried out with an immobilized pH gradient strip in the first dimension and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the second dimension. Over 800 polypeptide spots were resolved with a silver-staining protocol by computerized 2-D gel analsis. Seven of the polypeptide spots were evidently distinguishable between human left and right temporal lobes. Four of the polypeptide spots were larger and three were smaller in human right temporal lobe. One of these three protein spots that have descendent expression in human right temporal lobe was identified as carbonyl reductase (NADPH) 1 by MALDI-TOF MS. Thirty-three common spots were identified by ESI-MS/MALDI-TOF MS/Edman sequencing and a protein database search. These identified proteins include some important enzymes and regulating proteins. 相似文献