流感快检法与病毒分离法检测结果的比较

实验中用流感快检法和病毒分离法同时对61份标本进行检测,并进行了比较,快检法的阳性预测值为100%,阴性预测值为40.9%,两种方法的符合率为57.3%。结果表明,快检方法可以作为快速的辅助手段。     

一种快速微量提取植物叶片DNA的方法

介绍了一种快速提取微量DNA的方法。该方法简单易行,无需任何特殊设备,所需样品量少。提取的DNA纯度高,D260nm/D280nm在1．9-2．1之间,可满足RAPD、SSR、转基因植株的PCR检测等以PCR扩增为基础的实验需要。     

Rapid fixation and embedding for electron microscopy

A method has been developed for rapid processing of animal tissues for electron microscopy. The whole process of fixation staining dehydration, infiltration and embedding including polymerization is completed in less than 4 hr. A variety of human and animal tissues such as liver, spleen, muscle, kidney and embryonic chick heart were processed by this method and the results were excellent. The rapid fixation and embedding method is strongly recommended when relatively soft tissues are to be studied. This method is especially useful for examining pathological tissues for rapid diagnostic purposes.     

Application and efficiency of scintillation autoradiography for Drosophila polytene chromosomes

Summary A rapid method of autoradiography using the scintillation cocktail (Toluene and scintillation fluid, Omnifluor) has been described earlier. Its application and efficiency have been tested using both ³H-thymidine and ³H-uridine. The optimum time required for processing the autoradiograms has been found to be 24 h dry exposure followed by 48 h in the scintillation mixture. Detailed analysis of the autoradiograms with ³H-uridine reveals that with the rapid method the 100% level of labelling index is reached by 48 h while with the conventional method the same level is reached by 10 to 12 days of dry exposure. The maximum grain density is reached by 16 to 17 days by the conventional method. While by the rapid method, the maximum grain density is approximately 80% of the control, this grain density is reached by 48 h (plus 24 h of dry exposure) and thereafter forms a plateau. With Toluene alone the grain density never exceeds 20%. The background is also relatively low and less variable in the O-T-processed autoradiograms, as compared to the two controls. These results support that the scintillation fluid plays the key role in augmenting the labelling. Furthermore, although the maximum grain density by the rapid technique is 80% of the control, the grain density obtained by the rapid method gives less coincidence and superimposition of grains.On the other hand, with ³H-thymidine, although all labelling patterns could be resolved, the labelling index (i.e., percent of labelled cells) is about 40% at 48 h (plus 24 h) and about 79.5% at 96 h with the rapid method, as compared to about 30% and 44% with the conventional method at the two time points, respectively. Only with 16–17 days' dry exposure the ³H-thymidine labelling index increases to 67%. The frequency of the initial patterns (DD-2C) which are usually less frequent, has been found to have increased with the rapid method. No difference in grain density of labelling of ³H-thymidine could be detected between the rapid method and the conventional method. The resolution of grains also seems to be better by the rapid method, due probably to smaller size and lack of superimposition of grains. Other applications, advantages and limitations have been discussed.     

转基因植物快速检测方法的研究

本试验对转基因植物检测中的DNA提取和PCR扩增程序作了改进。经试验,本研究建立的DNA快速提取法与目前广泛使用的CTAB法相比更为简便,快速和经济,提取的DNA质量主扩增效果无明显差异,可用于多种转基因植物,多种植物组织的DNA提取,利用复合PCR法可在同一反应管中同步检测35N,NOS及CP4－EPSPS基因,明显提高了检测效率。应用本试验建立的DNA快速提取－复合PCR扩增－银染检测技术可在6小时内得出结果,达到了快速,简便,灵敏,可靠的检测目的。     

A rapid procedure for cell colony hybridization using DNA probes

A rapid, direct method for screening single cell-derived colonies or foci is described. The method allows the screening of a large number of colonies or foci by nitrocellulose filter hybridization using DNA probes. This technique simplifies current screening procedures and is a reliable, rapid, and sensitive method for the selection of cell clones containing a desired transfected gene.     

电化学法快速检测微生物的发展现状及趋势

自1898年Stewart提出利用电化学法检测微生物,电化学法已发展成为一种微生物快速检测的方法。根据检测的参数不同,电化学微生物检测法可以分为阻抗微生物法和介电常数法。阻抗法主要用于食品工业中微生物的快速检测(≤10⁷ cfu/mL),尤其用于易腐食品的微生物快速检测,以期实现在其发生明显腐败之前得到检测结果。而介电常数则用于生物发酵过程中的微生物数量的快速测定,可以实现在线监测微生物数量及生物发酵过程的实时控制。电化学法由于其检测迅速、可以实现自动化检测,在工业化生产中具有广阔的应用前景。     

一种供RAPD分析用蜘蛛模板DNA的快速提取方法

简要介绍了一种高效、快速提取蜘蛛模板 Ｄ Ｎ Ａ 供 Ｒ Ａ Ｐ Ｄ 分析的方法⒚该方法具有快速简便、重复性好的优点,提取的 Ｄ Ｎ Ａ 无污染物产生,完全能满足 Ｒ Ａ Ｐ Ｄ 研究的需要⒚     

A Review of Rapid Methods for the Analysis of Mycotoxins

An overview is presented of the analysis of mycotoxins by rapid methods such as: enzyme linked immuno-sorbent assay (ELISA); flow through membrane based immunoassay; immunochromatographic assay; fluorometric assay with immunoaffinity clean-up column or with a solid phase extraction clean-up column; and fluorescence polarization method. These methods are currently commercially available and are reliable, rapid methods. This review focuses on the basic principle of each rapid method as well as advantages and limitations of each method. Additionally, we address other emerging technologies of potential application in the analysis of mycotoxins.     

Rapid Fluorescent-Antibody Stain Technique with Group A Streptococci

A rapid fluorescent-antibody stain technique used with young broth cultures of trypsinized group A streptococci is described. Results from 1,214 trypsinized cultures of group A streptococci employing the rapid stain method were equivalent, both in specificity and sensitivity, when compared to results obtained from identical non-trypsinized cultures stained with the longer standard technique of Cherry, Goldman, and Carski. Moreover, the rapid method requires less than 2 min to complete.     

快速检测中药材乌梢蛇LAMP方法的建立

乌梢蛇作为一种名贵中药材,市面上伪品较多,干燥熏黑处理后的样品,更是真伪难辨。本研究致力于开发一套基于环介导等温扩增(loop-mediated isothermal amplification,LAMP)为基础的快速筛查乌梢蛇的方法。本研究以乌梢蛇12srRNA基因序列为基础设计并筛选出1套LAMP引物。通过调整反应条件,建立了对乌梢蛇LAMP的检查方法。结果显示,62℃下连续反应15min左右出现典型的"S"型荧光吸收曲线,实现了对乌梢蛇12srRNA基因序列的特异扩增。根据LAMP灵敏度高的特点,本研究简化了DNA的提取方法,缩短了检测的时间。相对于常规的PCR方法,本研究建立的以快速DNA提取为基础的乌梢蛇LAMP快速筛查方法具有简单、快速、灵敏、对设备要求低等特点,适用于对中药材乌梢蛇的快速筛查。     

A method to determine the mode of binding for GCPII inhibitors using bio-layer interferometry

The rapid dilution of the enzyme-inhibitor complex assay to monitor the recovery of enzyme activity is a well-established assay to determine the reversibility of inhibition. Our laboratory has previously employed this method to ascertain the reversibility of known glutamate carboxypeptidase II (GCPII)-targeting agents. Due to the tedious and time-consuming nature of the assay, we sought to develop a facile method to determine the reversibility of well-characterized GCPII inhibitors using bio-layer interferometry (BLI). The results from the BLI assay are in agreement with the rapid dilution method. Herein, we report for the first time, a rapid, novel real-time BLI method to determine reversibility of inhibition.     

一种适于PCR扩增的小麦基因组DNA快速提取法

许多小麦分子生物学研究需要对大量的小麦样品进行PCR检测,因此,建立一种快速提取小麦基因组DNA的方法十分必要。根据国外报道的一种快速提取水稻和玉米基因组DNA的方法,我们对部分提取步骤进行变动后,在小麦上进行了尝试,长度为1．5kb的片段能得到稳定的扩增。该方法样品研磨在1.5ml的离心管内进行,后续操作不用酚、氯仿、CTAB、SDS和巯基乙醇,整个提取过程不需要使用通风橱,操作步骤简单,花费时间少,而且提取的小麦基因组DNA完整性好,量也较可观。一个DNA样品可供50～100次PCR反应使用,适用于小麦遗传多样性、分子标记辅助选择、转基因后代检测以及引物筛选、分子标记定位等多种研究。     

水稻基因组DNA微量提取

随着植物学向分子水平的深入发展,研究中经常需要获得高质量的植物DNA样品,因此,建立植物DNA提取与纯化的常规实验方法对教学和科研都显得非常必要。介绍一种快速提取微量DNA的方法,该方法简单易行,无需任何特殊设备,所需样品量少,提取的DNA纯度高,可满足以PCR扩增为基础的实验需要。     

Rapid flow cytofluorometric analysis of mammalian cell cycle by propidium iodide staining

A rapid method for the flow microfluorometric determination of the DNA content per cell is described. Incubation of cells in a hypotonic solution of propidium iodide results in disruption of the cell membrane and rapid staining of nuclear chromatin. DNA distribution histograms generated from cells stained by this method are identical to those generated after fixation and RNase digestion. In contrast to some earlier described methods, the present technique is rapid (5 min of processing), requires a minimal amount of material, and avoids formation of cell clumps.     

Quality assurance in cervical smears: 100% rapid rescreening vs. 10% random rescreening

OBJECTIVE: To compare 100% rapid rescreening of cervical smears with 10% random rescreening as a method of quality assurance. STUDY DESIGN: A total of 5215 smears, randomly selected from smears reported as negative by cytotechnologists during routine screening, underwent 100% rapid rescreening by senior cytotechnologists. Ten percent of these smears, selected at random, were rescreened by other senior cytotechnologists. The gold standard was defined by cytopathologists, who rescreened all 5215 smears. After excluding unsatisfactory smears detected by cytopathologists, 4271 were included in the analysis. RESULTS: The 100% rapid rescreening method identified 69.9%, 95.7% and 100%, respectively, of atypical squamous cells of undetermined significance, low grade squamous intraepithelial lesion and high grade squamous intraepithelial lesion cases reported by the cytopathologists. The 100% rapid rescreening method showed a sensitivity of 73.5% and specificity of 98.6%. The 10% rescreening method showed sensitivity of 40.9% and specificity of 98.8%. CONCLUSION: One hundred percent rapid rescreening is an efficient method of internal quality assurance in cervical smear diagnosis. It can reduce the false negative rate and therefore can provide greater certainty to women who have received negative results. Well-trained cytotechnologists are able to identify abnormal smears in 1-minute rapid rescreening.     

A simplification of the protein assay method of Lowry et al. which is more generally applicable

Some recent modifications of the protein assay by the method of Lowry, Rosebrough, Farr, and Randall (1951, J. Biol. Chem.193, 265–275) have been reexamined and altered to provide a consolidated method which is simple, rapid, objective, and more generally applicable. A DOC-TCA protein precipitation technique provides for rapid quantitative recovery of soluble and membrane proteins from interfering substances even in very dilute solutions (< 1 μg/ml of protein). SDS is added to alleviate possible nonionic and cationic detergent and lipid interferences, and to provide mild conditions for rapid denaturation of membrane and proteolipid proteins. A simple method based on a linear log-log protein standard curve is presented to permit rapid and totally objective protein analysis using small programmable calculators. The new modification compared favorably with the original method of Lowry et al.     

线粒体DNA高变区hν1快速测序方法及应用

本研究目的是使线粒体DNAhv1区的测序更加简单、快速和准确,以利于法医学检案。将线粒体DNAhv1高变区分别用三对重叠的引物进行全序列扩增,对这三对引物的扩增产物进行测序分析。用经过大量实验反复摸索得到的稳定的实验条件,对已确定为同一母亲所生的兄弟二人的样品进行分析,结果完全一样;对待确定姐弟关系的样品进行分析,结果是认定的;操作时间较原方法缩短一半。结果表明该方法是可替代原法的快速测序法。     

Automated rapid method for microbioassay of amino acids

An automated rapid method for microbioassay of amino acids was investigated by taking advantages of the rapid manual method. The Pye Unicam automatic analytical apparatus was adopted for the automation of assay culture in the rapid microbioassay. The advantages of the present method are that amino acids can be automatically determined on 3.5-hr assay culture, and that an aseptic technique can be omitted. These advantages were confirmed in several amino acid assays. The assay values were the same as those obtained by the conventional method. Application of the automated rapid method to the serine assay showed a linear standard curve without a lag section, leading to more expanded assay range and smaller drift in values.