共查询到20条相似文献，搜索用时 531 毫秒
1.
目的 评价和分析一种新的人类K-ras基因突变检测试剂盒质量,确定该类试剂质量标准和评价方法.方法 利用实时荧光定量PCR方法,检测7种K-ras基因单一突变点质控品的准确性、特异性、最低检测量和重复性.分别检测7例肠癌患者K-ras基因突变阳性组织样本和10例阴性样本的准确性和特异性 .结果 试剂盒准确性、特异性、最低检出量和重复性均符合质量标准.能准确和特异的检出单一点突变,具有较好的灵敏度和重复性.结论 该试剂盒质量标准设定较合理,在指导K-ras基因突变检测方面具有较好的应用前景. 相似文献
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荧光探针是科学研究的一种新型工具,具有特异性强、灵敏度高和生物相容性好等特点,应用于众多生产生活领域。介绍了荧光探针的结构、分类及反应机理,综述了荧光探针在食品领域及生物学领域中的应用研究进展。主要从食品重金属检测、食物新鲜度检测、细胞活度检测、细胞内活性物质检测等方面介绍了荧光探针的应用实例,并展望其发展前景,旨在为今后荧光探针类高附加值产品的开发与设计提供可行性参考。 相似文献
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设计、合成了一种基于罗丹明B的铜离子荧光分子探针RF。研究发现在V乙腈∶V水=1∶1介质中,化合物RF最大发射波长为566 nm,Cu2＋可使探针RF的荧光强度明显增大,同时溶液颜色从无色变为桃红色。该荧光探针对Cu2＋具有较高的灵敏度和较好的选择性。RF的选择性荧光增强是由于其对Cu2＋的识别开环引起的。该探针对Cu2＋的识别是不可逆的,而且pH对探针RF在中性水溶液中检测Cu2＋几乎没有影响。 相似文献
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王志平 《渭南师范学院学报》2012,(10):61-64
荧光定量PCR技术是在普通PCR技术基础上建立起来的一种利用标准曲线对未知模板进行定量分析的方法.它特异性强、灵敏度高、操作简便、快速高效,在疾病诊断、食品检测、环境监测以及生物学研究等方面具有广泛的应用前景.在阐述荧光定量PCR技术原理的基础上,系统论述了该技术在各方面的应用. 相似文献
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目的:介绍一种快速简便地优化多重PCR检测DMD基因外显子缺失的方法及详细步骤.方法:采2ml外周血,用0.2%氯化钠处理收集白细胞,再用基因组DNA提取试剂盒抽提基因组DNA,用优化的多重PCR法直接检测DMD基因外显子的缺失.结果:用该方法检测DMD基因外显子缺失结果准确清晰.结论:用优化的多重PCR技术可以直接检测DMD基因外显子缺失,跟通常使用的9对引物一步法相比,具有经济、快速、简便等特点. 相似文献
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质粒DNA提取方法的比较 总被引：1，自引：0，他引：1
以含有原核表达载体pet21a和真核表达载体EGFP的DH5α系列菌株为材料,分别采用碱裂解法和试剂盒提取法提取质粒DNA,通过凝胶电泳分析,对两种质粒提取方法的各自特点进行了比较与分析。认为碱裂解法和试剂盒提取法提取质粒DNA均可进行后续试验,但试剂盒提取法具有省时、简洁、高效的优势。同时根据在实验过程中所遇到的问题,说明了在进行质粒DNA提取过程中应当注意的问题。 相似文献
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依托教师的科研成果,设计了"超顺磁性纳米簇捕获和分离细菌综合性实验".利用高温水解法制得超顺磁性纳米簇,进一步修饰沙门氏菌抗体,构建磁性纳米探针,表征了其形貌、磁性和生物活性,并详细探究了对沙门氏菌的分离效率、特异性及抗干扰能力.实验结果表明:磁性纳米探针能够快速、特异、高效地捕获和分离沙门氏菌,捕获效率达95％以上,... 相似文献
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《赣南师范学院学报》2019,(3):69-72
开发简单快速的汞离子(Hg~(2+))检测新方法对人类了解重金属污染具有重要的意义.基于G-三链体DNA探针设计一种非标记的荧光方法用于汞离子检测.富含G碱基的DNA序列能自主装形成G-三链体结构.在没有Hg~(2+)存在的情况下,G-三链体DNA能与荧光染料硫黄素T结合生成非标记的荧光探针.硫黄素T与G-三链体结合后,ThT的荧光信号会显著增强.在Hg~(2+)存在的情况下,G-三链体DNA环部的胸腺嘧啶T能与Hg~(2+)发生反应,从而改变DNA的构型,抑制G-三链体结构的形成,使得ThT的荧光减弱.该方法通过对ThT-G-三链体的荧光强度变化实现溶液中的Hg~(2+)离子的检测.对该方法的可行性进行论证,并对实验条件进行优化.实验结果表明该方法可用于Hg~(2+)检测,并具有良好的灵敏度和选择性. 相似文献
9.
实时荧光定量PCR技术的操作实践 总被引：5，自引：0，他引：5
采用Mx3000P型实时荧光定量PCR仪,以SYBR(R) Green I法相对定量技术为例,设计1例检测针对HIV-1 vpr基因的特异性siRNA基因沉默效果的实验.介绍了实时荧光定量PCR技术上机前样本的制备过程、检测程序及相关参数的设置方法等操作流程;结合熔解曲线、扩增曲线进行结果分析;强调了实验整体设计、引物设计与PCR反应体系的优化和内参的恰当选择在实时荧光定量PCR技术中的重要性. 相似文献
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食品沙门氏菌PCR快速检测试剂盒简介 总被引：6，自引：0，他引：6
在我国食源性疾病中细菌性食物中毒病例最为常见，其中由沙门氏菌引起的中毒病例占70％-80％，而以肉、蛋、奶等畜产品被沙门氏菌污染所引起的中毒可占90％以上。因此，目前对食品质量安全监控进行CMA认证检测时，沙门氏菌的检测已成为了必检的卫生指标测定项目之一。传统的检测方法过程复杂，周期长，不仅造成人力物力的浪费，也影响了口岸的通关速度。 相似文献
12.
合成了对铜离子有很强选择性的罗丹明类荧光探针。在乙腈/水（体积比1∶1）溶液中,当加入Cu2＋后,探针显桃红色,随Cu2＋浓度增大,荧光强度增强,发射荧光波长红移。并在2.8×10-7~2.8×10-5mol/L范围内,Cu2＋离子浓度与荧光强度呈现良好的线性关系。其它常见离子引起很小的荧光光谱变化,合成的试剂可用于高选择性的检测铜。 相似文献
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以1,8-萘二甲酸酐、二乙烯三胺（DETA）、N-甲基哌嗪及丙烯基氯为原料,通过酰胺化、季胺化、SN2亲核取代以及丙烯酰胺共聚合等反应,合成了水溶性1,8-萘酰亚胺高分子荧光探针。用紫外光谱、荧光光谱等手段研究它们在水、四氢呋喃和乙醇溶液中光物理化学性质,同时考察浓度和溶剂极性及取代基对荧光性能影响及对金属离子的识别作用。结果表明,此高分子荧光探针的光稳定性及荧光量子产率明显提高,随着溶剂极性增大,荧光量子产率增大,波长红移;当浓度超过8×10^-4g/m L时出现荧光浓度自猝灭;该探针在水中能对Cu^2＋在392 nm处进行高选择性识别。 相似文献
14.
The clinical diagnosis of sepsis is difficult, particularly in neonates. To devise a rapid and reliable method for identifing
bacteria in blood and cerebrospinal fluid (CSF), we developed a pair of primers according to the gene encoding 16 s rRNA,
found in all bacteria. DNA fragments from different bacterial species and from clinical samples were detected with polymerase
chain reaction (PCR), and with reverse hybridization using a universal bacterial probe, a gram-positive probe and a gram-negative
probe. Our results showed that a 371 bp DNA fragment was amplified from 20 different bacterial species. No signal was observed
when human DNA and viruses were used as templates. The sensitivity could be improved to 10−12 g. All 26 culture-positive clinical samples (22 blood samples and 4 CSF samples), were positive with PCR. The gram-negative
and gram-positive probe hybridized to clinical samples and to known bacterial controls, as predicted by Gram’s stain characteristics.
Our results suggest that the method of PCR and reverse hybridization is rapid, sensitive and specific in detecting bacterial
infections. This finding may be significant in the clinical diagnosis of sepsis in neonates.
Project (396457) supported by the Zhejiang Provincal Natural Science Foundation. 相似文献
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Wang JY Gu YS Wang J Tong Y Wang Y Shao JB Qi M 《Journal of Zhejiang University. Science. B》2008,9(8):610-615
Objective:Leber's hereditary optic neuropathY (LHON)is a maternally inherited degeneration of the optic nerve caused by point mutations of mitochondrial DNA(mtDNA).Many unsolved questions regarding the penetrance and pathophysiological mechanism of LHON demand efficient and reliable mutation testing.This study aims to develop a minor groove binder(MGB) probe assay for rapid detection of mtDNA11778 mutation and heteroplasmy in Chinese LHON patients by real-time polymerase chain reaction(PCR).Methods:Forty-eight patients suspected of having LHON and their maternal relatives underwent a molecular genetic evaluation,with 20 normal individuals as a control group at the same time.A real-time PCR involving two MGB probes was used to detect the mtDNA 11778 mutation and heteroplasmy.A linear standard curve was obtained by pUCmLHONG and pUCmLHONA clones.Results:All 48 LHON patients and their matemal relatives were positive for mtDNA 11778 mutation in our assay,27 heteroplasmic and 21 homoplasmic.Eighteen cases did not show an occurrence of the disease,while 9 developed the disease among the 27 heteroplasmic mutation cases.Eleven did not show an occurrence of the disease,while 10 cases developed the disease among 21 homoplasmic mutation cases.There was a significant difierence in the incidence between the heteroplasmic and the homoplasmic mutation types.The time needed for running a real-time PCR assay was only 80 min.Conclusion:This real-time PCR assay is a rapid,reliable method for mtDNA mutation detection as well as heteroplasmy quantification.Detecting this ratio is very important for predicting phenotypic expression of unaffected carriers. 相似文献
16.
阐述超声波A扫探伤过程中焊缝缺陷波动态波形特点与探头的检测方式有关,指出识别出探头的检测方式对缺陷的智能识别至关重要。利用Matlab工具箱中的图像处理模块对超声波检测中探头位置识别,并确定出探头的检测轨迹。实践表明,该方法具有一定的快速性和实用性。 相似文献
17.
以CdTe量子点为荧光探针,基于荧光猝灭法对Ag（Ⅰ）和Ca（Ⅱ）进行了定量检测.考察了缓冲液的浓度、缓冲液pH值、反应时间等多种因素的影响.结果表明：（1）在浓度为10～20 mmol/L、pH值为7～8的磷酸二氢钠-磷酸氢二钠缓冲液中,Ag＋与量子点反应时间为20 min时,量子点荧光衰减程度（ΔF）与Ag＋浓度呈... 相似文献
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Detection of PCV2 DNA by SYBR Green I-based quantitative PCR 总被引：5，自引：0，他引：5
We developed an assay for the detection and quantitation of porcine circovirus type 2 (PCV2) with the SYBR Green I-based real-time PCR. The real-time PCR provides a broad dynamic range, detecting from 103 to 1011 copies of DNA per reaction. No cross-reactions were found in specimens containing PCV1. Because of the high sensitivity and specificity of the assay with a relatively rapid and simple procedure, real-time PCR can be used as a routine assay for the clinical diagnosis of PCV2 infection. In this study we applied real-time PCR assay to 80 clinical samples, collected from 40 pigs with postweaning multisystemic wasting syndrome (PMWS) and 40 healthy pigs in comparison with conventional PCR assay. In 56 of 80 samples, PCV2 DNA was de-tected by conventional PCR assay. All samples positive for PCV2 DNA in conventional PCR assay were also positive in real-time assay, and 12 of 24 samples that tested negative for PCV2 DNA in the conventional assay were tested positive in real-time PCR assay. Real-time PCR assay increased the number of samples in which PCV2 was detected by 15%. It is, therefore, considered to be a useful tool for the detection of PCV2. 相似文献
20.
In the search for a rapid and reliable method for identification of bacteria in blood and cerebrospinal fluid , we developed a unified set of primers and used them under polymerase chain reaction(PCR) to amplify the spacer regions between the 16s and 23s genes in the prokaryotic rRNA genetic loci . Spacer regions within these loci showed a significant level of length and sequence polymorphism across most of the species lines. A generic pair of priming sequences was selected from highly conserved sequences in the 16s and 23s genes occurring adjacent to these polymorphic regions. This single set of primers and reaction conditions were used for the amplification of the 16s-23s spacer regions for 61 strains of standard bacteria and corresponding clinical isolates belonging to 20 genera and 27 species, including Listeria, Staphylococcus and Salmonella species, et al. When the spacer amplification products were resolved by electrophoresis, the resulting patterns could be used to distinguish most of the bacteria species within the test group, and the amplification products of the clinical isolates clustered at the standard species level. Some species presenting similar pattern were further analyzed by HinfI or AluI digestion or DNA clone and sequences analysis in order to establish the specific 16s-23s rRNA gene spacer regions map. Analysis of 42 blood specimens from septicemic neonates and 6 CSF specimens from suspected purulent meningitis patients by bacterial culture and PCR-RFLP(Restriction Fregament Length Polymorphism) showed that 15 specimens of blood culture were positive(35.7%) in the 42 septicemic neonates; 27 specimens were positive(64.2%) by PCR, and that the positive rate by PCR was significantly higher than that by blood culture(P<0.01). Among the 6 CSF specimens, one specimen found positive by blood culture was also positive by PCR, two found negative by blood culture showed positive by PCR; all three were S.epidermidis according to the DNA map. One C.neoformans found positive by blood culture showed negative by PCR. The remaining two specimens were both negative by PCR and blood culture. These results indicated that the method of detecting bacterial 16s-23s rRNA spacer regions using PCR and RFLP techniques was rapid, sensitive and specific in the detection of bacterial infections; and so, has very important application in the clinical diagnosis of sepsis in neonates. 相似文献