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排序方式: 共有148条查询结果,搜索用时 15 毫秒
1.
本研究旨在建立食品中产呕吐毒素蜡样芽胞杆菌的快速检测方法。基于产呕吐毒素蜡样芽胞杆菌合成酶基因cesB靶基因,设计4条特异性引物(2条内引物、2条外引物),建立环介导等温扩增技术(LAMP)。然后采用2株产呕吐毒素蜡样芽胞杆菌、19株蜡样芽胞杆菌和41株非蜡样芽胞杆菌验证了该LAMP具有很好的特异性。LAMP检测的灵敏度在DNA水平上达到1.49 pg/μL,纯菌的灵敏度为5×10~3 cfu/mL。人工污染米饭样品,当起始污染量为2 cfu/g时,37℃增菌培养6 h,用试剂盒法和煮沸法提取的DNA,都可以检出产呕吐毒素蜡样芽胞杆菌。采用此LAMP方法进行了72份实际样品的检测,检出2份产呕吐毒素蜡样芽胞杆菌阳性样品,与传统方法一致。因此,本研究建立的LAMP检测方法可应用于食品中产呕吐毒素蜡样芽胞杆菌的快速特异检测。  相似文献   
2.
嵌入式Web控制终端的物联网移动E-Laboratory平台   总被引:1,自引:0,他引:1       下载免费PDF全文
为了进一步提高大学校园设备资源的利用率,实现多种实验装置的网络化控制和用户终端移动化,基于物联网技术,将LAMP移植到采用ARM架构的微处理器上,通过搭建嵌入式Web控制终端,对Web服务平台进行了详细的设计,实现了多种实验装置的网络化控制;将单自由度机械臂系统这一实验装置与EWC_T连接,基于Android操作系统上开发了一个应用,设计并搭建了Mobile Experiment系统,实现了网络化实验室平台的用户终端的移动化;通过实验证明该系统能够可靠地完成对实验装置的远程控制,并具有低功耗、低成本和便携性好等优点。  相似文献   
3.
目的建立环介导等温扩增法(loop-mediated isothermal amplification,LAMP)检测酸奶中嗜酸乳杆菌。方法以嗜酸乳杆菌16S r RNA保守区的基因序列为靶序列设计引物,优化建立LAMP反应体系和程序,并研究引物的特异性,同时确定嗜酸乳杆菌的灵敏度和检出限。结果本方法可通过凝胶电泳和离心后焦磷酸镁白色沉淀检测反应结果,检测时间为5.5 h,灵敏度为62 CFU/m L,人工污染酸奶样品的检出限为120 CFU/m L,12株非嗜酸乳杆菌细菌非特异检测结果均为100%。结论该方法灵敏性好、特异性强、操作简单、耗时短,可适合于酸奶中的嗜酸乳杆菌的快速测定。  相似文献   
4.
建立了一个鉴定牛羊肉中搀杂杂动物肉的环介导恒温扩增检测方法。确定了一套可在牛羊肉中特异并灵敏地检测出搀杂肉成分的引物对,以动物细胞色素b基因组为模板可在恒温63℃恒温特异性扩增出猪等基因片段而无其他扩增片段影响。可检测牛肉中0.01%~2%的猪肉成分。经与PCR方法比对,结果表明,该方法可有效用于实际生鲜肉或加工肉制品样本的鉴定。  相似文献   
5.
Loop‐mediated isothermal amplification (LAMP) is a novel method that amplifies target nucleic acids under isothermal conditions. It is a rapid, specific, and sensitive method, which does not require costly thermal cyclers for the detection of nucleic acids. Thus, it is suitable for on‐site detection assays under low‐resource settings. It can also be integrated on compact lab‐on‐a‐chip devices for the development of micro‐total analysis systems. This review discusses LAMP‐based methods, as well as LAMP‐based centrifugal, microfluidic, and other fluid‐handling devices, which have been developed for the assessment of meat quality parameters that are related to the presence or absence of nucleic acids, for example, animal species identification and microbiological quality. Advances in improving the rapidity, specificity, and sensitivity of LAMP techniques for the assessment of these meat quality parameters are also discussed in this review.  相似文献   
6.
建立食品中铜绿假单胞菌环介导等温扩增(LAMP)检测方法。利用铜绿假单胞菌外毒素A(PEA)基因序列,设计3对铜绿假单胞菌LAMP检测特异性引物,用36株铜绿假单胞菌,14株近源菌验证方法的特异性。建立的LAMP方法特异性好,灵敏度达到2.2 cfu/100 g~3.5 cfu/100 g。建立的食品中铜绿假单胞菌LAMP检测方法特异性好,灵敏度高,适用于的检测食品中的铜绿假单胞菌。  相似文献   
7.
We evaluated the usefulness of the loop-mediated isothermal amplification (LAMP) using a portable ESE Quant tube scanner as a rapid and simple method for the detection of Vibrio parahaemolyticus, an important pathogen causing seafood-borne gastroenteritis. The real time LAMP (RT-LAMP) assay using a hemolysin gene (tlh/ldh)-specific primers was verified using V. parahaemolyticus strains (n = 91) from different countries and other non-target strains to check the utility of this method. Both the sensitivity and specificity of the RT-LAMP using 3 pairs of tlh/ldh-specific primers developed in this study were excellent (100%). The detection limit of the RT-LAMP was as low as 7 Colony forming unit per reaction and detection time was only 20 min. Comparative evaluation of the target bacterial strains with the RT-LAMP using ESE Quant tube scanner, API 20E system and conventional RT-PCR method revealed that the RT-LAMP assay developed in this study is simpler and more rapid than the latter two methods. Therefore, the RT-LAMP method using the easily portable ESE Quant tube scanner can be considered as an effective tool for the rapid screening of V. parahaemolyticus strains in environmental and clinical samples, especially, in remote areas of developing countries during epidemic periods.  相似文献   
8.
陈文秀  姜旋  马晓燕  张伟 《食品科学》2014,35(20):192-197
为了建立食品中肺炎克雷伯氏菌灵敏快速的检测方法,根据GenBank中已公布的肺炎克雷伯氏菌phoE基因设计引物,利用实时荧光监测仪,建立实时荧光环介导等温扩增(loop-mediated isothermal amplification,LAMP)技术检测食品中肺炎克雷伯氏菌的方法,可直观判定检测结果,仪器亦可自动显示检测结果。对该检测方法的特异性、灵敏度等方面进行研究,并与聚合酶链反应(polymerase chain reaction,PCR)检测方法进行比较。结果表明,实时荧光LAMP检测方法特异性强、灵敏度高。用于特异性实验的21株实验菌株中,4株肺炎克雷伯氏菌,呈现阳性结果,而其他17株非肺炎克雷伯氏菌,均呈阴性结果;检测纯菌的灵敏度和检测人工污染乳粉的检出限分别为79 CFU/mL和79 CFU/g,是常规PCR检测方法的100倍。总之,本研究建立的实时荧光LAMP检测方法灵敏度高、特异性强、耗时短、结果判定直观,实现了对肺炎克雷伯氏菌的快速检测。  相似文献   
9.
刘津  张隽  李婷  张璜  高东微  李志勇 《食品科学》2014,35(22):226-232
根据澳洲坚果豌豆蛋白AMP2基因序列,利用设计软件Primer Explorer Version 4设计并筛选了食品过敏原澳洲坚果的环介导等温扩增引物,对反应体系和反应条件进行优化,建立澳洲坚果的环介导等温扩增检测方法,结果判断可采用实时荧光法和荧光染料终点显色法。对该方法进行了特异性、灵敏度、稳定性评价,结果显示:该方法能够特异性、灵敏、稳定地检测食品中的澳洲坚果成分,检测低限为0.5%。此外,对7 种市售食品样品的检测结果表明,该方法与食品标签标示的过敏原成分结果吻合率为100%,假阳性率和假阴性率均为0,在市售食品的过敏原成分检测上较商业化快速检测试纸条更加稳定可靠。  相似文献   
10.
Vibrio parahaemolyticus is a crucial foodborne pathogen. The viable but non-culturable (VBNC) state of V. parahaemolyticus cannot be detected by traditional culture methods. The objective of this study was to develop and evaluate a method that combines propidium monoazide (PMA) treatment with real-time fluorescent loop-mediated isothermal amplification (RF-LAMP) to detect and quantify VBNC cells of V. parahaemolyticus. Different states of cells were treated with PMA in dark for 5 min and subsequently exposed to a 650 W halogen lamp for 5 min. The cells were collected and DNA was amplified by RF-LAMP. The primers which targeted six distinct regions in the tlh gene of V. parahaemolyticus were used for the PMA-RF-LAMP assay. The results indicated that the treatment with 4 μg/mL of PMA and a further exposure to light for 5 min was suitable for PMA-RF-LAMP to distinguish viable cells from dead cells of V. parahaemolyticus. The developed assay exhibited remarkably high specificity, sensitivity and rapidity, without any cross reaction with the tested non-V. parahaemolyticus strains. There was good linear correlation between Ct values and log copy/mL in the range of 6.8–6.8 × 107 copy/mL, with the reaction endpoints less than 30 cycles (30 min). It could detect as low as 14 copy/g of V. parahaemolyticus in spiked seafood samples (pomfret, shrimp, scallop, oyster and salted fish) without any interference from food matrices, dead cells and other bacteria.  相似文献   
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