Outbreak in Bristol of Conjunctivitis Caused by Adenovirus Type 8, and its Epidemiology and Control

In an investigation of an outbreak of adenovirus type 8 infection involving over 40 people it was found that the infection spread initially within the eye hospital, but subsequently involved several family contacts and two local hospitals for mentally subnormal patients. Presumptive diagnosis should be possible before subepithelial opacities have developed provided an adequate history is taken, and had that been done in this outbreak it is reasonable to suppose that many cases could have been prevented.     

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。     

食源性致病菌快速检测技术研究进展

食源性致病菌是影响食品安全的主要因素之一,传统的细菌分离、培养与鉴定由于需时较长,特别是有的细菌难以培养,难以适应食源性疾病预防控制的需要,因而快速、简便、特异的检测方法成为研究的热点。对电阻抗、放射测量、微热量、ELISA、PCR、基因芯片和生物传感器技术在金黄色葡萄球菌、沙门菌、肠出血性大肠埃希菌等食源性致病菌快速检测中的应用研究进行综述。     

硫解高效液相色谱法快速检测原花青素

为开发一种苄硫醇分解-高效液相色谱检测原花青素的新方法,本研究制备并纯化了苄硫醚-表儿茶素和苄硫醚-表儿茶素没食子酸酯,并与没食子酸、儿茶素、表儿茶素和表儿茶素没食子酸酯为对照品,在新的色谱条件下6种硫解产物可在11 min之内完成分离,外标法定量分析RSD均小于1.86%(n=5),最低检出限为0.5～1.5 ng,在1 ng～960 ng之间线性关系良好(r≥0.9996),平均回收率77.8%～94.6%。这种方法可用于植物及其提取物中原花青素含量及平均聚合度的快速分析。     

乳鼠脑组织中牛磺酸的快速检测

建立一种快速、准确的牛磺酸定量检测方法。采用Beckman公司6300黄金系统氨基酸分析仪,在锂柱130 min程序生理体液分析方法基础上,根据牛磺酸(TAU)的特性,建立了脑组织中TAU快速测定方法。用此方法完成TAU分析的时间为17 min,比原方法缩短了123 min。且有较好的重现性(日内RSD 0.42%,日间RSD 0.57%)、回收率高(98.39%)。本方法简便、快速、准确、可靠,适用于临床和科研工作。     

Rapid Detection of Certain Basic Drugs in Urine

Equipment costing only a few pounds can be used to detect a wide range of drugs in urine by anyone with a minimum of scientific knowledge. A run of tests takes about half an hour to perform and cost about 10p.     

猴腺病毒（SAdV）PCR检测方法的建立及初步应用

目的建立SAdV特异的PCR检测方法并研究实验猴群和猴源性生物制品中SAdV感染或污染情况。方法比对分析多株SAdV序列,设计SAdV特异引物,优化PCR实验条件,建立的PCR方法经验证后检测实验猴群和猴源性生物制品,阳性产物测序并构建进化树。结果经特异性和敏感性鉴定,在设计的5对引物中确定一对最佳引物,可以区分SAdV和MAD,ICH,CELO且可以检测到的最小DNA量为47.9 pg/mL。PCR方法检测实验猴群,阳性率49.2%,测序及进化分析表明,SAdV感染型别呈广泛基因多样性。主要分布在G亚属和以SAdV-49为代表的分支。结论经测序验证,PCR检测方法具有很好准确性,初步应用表明我国实验猴群中SAdV高度流行,应加强实验猴群及相关生物制品中SAdV的监测,避免人类感染SAdV的潜在风险。     

HPLC-ELSD快速测定微藻中性脂

溶剂法提取微藻油脂不同于植物种子油脂,它是全细胞的提取物,成分非常复杂,存在与甘油三酯(TAG)在色谱保留性质上相近的低极性物质,干扰TAG的测定。建立了基于二醇基柱及蒸发光检测器、正己烷-异丙醇为流动相的快速测定微藻中性脂的方法。对该方法进行评价,结果显示,测定的TAG、游离脂肪酸(FFA)、甘油二酯(DAG)、甘油一酯(MAG)线性相关系数均大于0.99,方法重复性好。湛江等鞭金藻及微拟球藻样品中TAG加标回收率为96.2%~113.1%,相对标准偏差(RSD)为0.46%~4.8%。将本方法测定湛江等鞭金藻及微拟球藻中TAG的含量并与传统的固相萃取(SPE)及常用的TLC/GC测定TAG的方法进行比较,相比上述两种方法,该方法前处理简单、灵敏度高,可快速准确测定微藻中TAG的含量。     

快速检测支气管肺泡灌洗液T细胞的研究

目的建立一种简便、快速检测支气管肺泡灌洗液（ＢＡＬＦ）中Ｔ淋巴细胞总数的方法。方法用改良的α－醋酸萘酯酶（ＡＮＡＥ）染色法鉴别和计数ＢＡＬＦ中Ｔ淋巴细胞。用本法共检测２２７例大鼠ＢＡＬＦ,并采用两样本均数比较的ｔ检验分析３批动物实验的测定结果。结果大鼠ＢＡＬＦ细胞涂片ＡＮＡＥ染色效果满意,Ｔ淋巴细胞容易鉴别。测得１１０只正常大鼠ＢＡＬＦ中Ｔ淋巴细胞比值的正常值为（６９．３６±１１．４７）％。各个实验组与相应对照组比较,ＢＡＬＦ中Ｔ淋巴细胞比值均出现不同程度的变化。结论本方法简便易行,能准确地显示ＢＡＬＦ中Ｔ淋巴细胞数值的变化。本研究是国内首次报道,为实验和临床研究肺局部细胞免疫状况提供了一种简便的方法及其应用的实验依据。     

实时荧光PCR快速检测食品中致敏原芹菜成分

探讨采用实时荧光PCK技术建立检测食品中致敏原芹菜成分的方法.试验结果表明:针对芹菜管家基因Mrd基因设计的特异性检测引物和探针进行检测时,31种样品经实时PCR扩增,只有芹菜和西芹样品产生阳性的荧光信号,其余样品均不产生荧光信号.灵敏度试验结果表明:该方法对芹菜致敏原基因的检测灵敏度较高,可检测到芹菜过敏原样品中10mg/kg～1mg/kg的含量.     

Rapid Methods for High-Throughput Detection of Sulfoxides

Enantiopure sulfoxides are prevalent in drugs and are useful chiral auxiliaries in organic synthesis. The biocatalytic enantioselective oxidation of prochiral sulfides is a direct and economical approach for the synthesis of optically pure sulfoxides. The selection of suitable biocatalysts requires rapid and reliable high-throughput screening methods. Here we present four different methods for detecting sulfoxides produced via whole-cell biocatalysis, three of which were exploited for high-throughput screening. Fluorescence detection based on the acid activation of omeprazole was utilized for high-throughput screening of mutant libraries of toluene monooxygenases, but no active variants have been discovered yet. The second method is based on the reduction of sulfoxides to sulfides, with the coupled release and measurement of iodine. The availability of solvent-resistant microtiter plates enabled us to modify the method to a high-throughput format. The third method, selective inhibition of horse liver alcohol dehydrogenase, was used to rapidly screen highly active and/or enantioselective variants at position V106 of toluene ortho-monooxygenase in a saturation mutagenesis library, using methyl-p-tolyl sulfide as the substrate. A success rate of 89% (i.e., 11% false positives) was obtained, and two new mutants were selected. The fourth method is based on the colorimetric detection of adrenochrome, a back-titration procedure which measures the concentration of the periodate-sensitive sulfide. Due to low sensitivity during whole-cell screening, this method was found to be useful only for determining the presence or absence of sulfoxide in the reaction. The methods described in the present work are simple and inexpensive and do not require special equipment.The growing demand for green catalytic processes has increased the utilization of enzymes as industrial biocatalysts for the synthesis of fine chemicals (6, 19, 20). As a consequence, there is a continuous search for novel or improved biocatalysts. In order to find an appropriate candidate for a process, various sources of enzymes must be screened for activity (23). Therefore, a sensitive, reproducible, accurate, and simple high-throughput screening method is a key prerequisite for the development of biocatalytic processes on an industrial scale (32, 39).Screening systems are divided into three different classes. The first class contains assays applicable to testing growing or resting microbial colonies for enzymatic activity directly on agar plates (23), for example, detection of epoxide hydrolase activity on butane oxide by use of safranin O. Oxidation of the 1,2-diol product by Escherichia coli modified the membrane potential and led to accumulation of the red dye in the colonies producing active enzyme (34). In another study, the spontaneous oxidation of substituted catechols to brown-red quinones was used to screen random libraries of whole cells expressing toluene monooxygenases (TMOs) for regioselective oxidation of substituted phenols (12, 30). The positive clones produced a red halo around the cells. These assays are high-throughput, simple procedures but often require a tailored substrate with a chromophore, such as bromonaphthol or azo-dye (23).The second class includes chromogenic and fluorogenic assays applicable in microtiter plates or microarray formats (23). Microtiter plates in 96- or 384-well format are particularly well suited for spectroscopic reading using either UV-visible or fluorescence plate readers. This class may be subdivided into the following four groups: (i) enzyme-coupled assays, such as the determination of dehydrogenase activity through formation of NADH from NAD and an absorbance change at a wavelength of 340 nm; (ii) assays using chromogenic and fluorogenic substrates, such as various synthetically labeled substrates that are commercially available for the determination of hydrolytic activity produced by lipases, phosphatases, glycosidases, amidases, etc.; (iii) assays using chromogenic and fluorogenic sensors, such as widely used pH indicators (16), that may be applied in any reaction that includes a change in pH; and (iv) microarray assays using a solid support, enabling screening of thousands of samples. The high-throughput potential of these methods was demonstrated by profiling of 40 different esterases and lipases across 35 different fluorogenic ester substrates, using only 50 μl of each enzyme solution and a submilligram quantity of each substrate for over 7,000 tests (2).The third class of enzymatic assays rely on product detection by instruments and include gas chromatography (GC), high-pressure liquid chromatography (HPLC), mass spectrometry, nuclear magnetic resonance (NMR) spectrometry, and infrared radiation assays that have been adapted for high throughput (22, 23, 33). Such assays require expensive and sophisticated equipment, but they allow working directly with the substrate of interest and are rapidly adapted once the instrument is available (23).Various chemical substances can be synthesized by bacteria and fungi, among which are the chiral sulfoxides (5, 10, 11, 24, 36). As natural products, chiral sulfoxides possess a wide range of biological activities, from flavor and aroma precursor activities to antimicrobial properties. In addition, they are efficient auxiliaries that lead to essential asymmetric transformations (3, 11). Furthermore, one of the most significant applications of chiral sulfoxides is in the pharmaceutical industry (3). The world''s best-selling antiulcer drug, (S)-omeprazole, is a chiral sulfoxide (11, 14). Although there have been numerous reports on chemical and biological methods for synthesizing chiral sulfoxides, little information exists about rapid high-throughput assays for sulfoxide determination. In this study, four colorimetric or fluorometric procedures were evaluated and adapted for screening of whole-cell libraries containing variants of TMOs. Three of the four methods were exploited successfully to a high-throughput format using 96-well microtiter plates, whereas one method was not suitable due to low sensitivity. The method based on acid activation of omeprazole proved very efficient, but no positive variants were found, whereas the one based on selective inhibition of horse liver alcohol dehydrogenase (HLADH), originally reported by Sprout and Seto (28), was useful for detecting mutants with high activity and enantioselectivity in the oxidation of methyl p-tolyl sulfide.     

Method for Rapid Detection of Cyanogenic Bacteria

An agar plate method is described in which the production of hydrogen cyanide by as many as 50 microbial isolates per plate may be detected. Cyanide produced by the organisms reacts with copper(II) ethylacetoacetate and 4,4′-methylenebis-(N,N-dimethylaniline) in a paper disk suspended above the microbial colonies. Cell growth occurs in depressions in the agar surface, which allows separation of colonies and enhances sensitivity of hydrogen cyanide detection.     

食品中金黄色葡萄球菌肠毒素的快速检测方法

采用反向被动乳胶凝集法 (RPLA)和mini VIDAS (ELFA)两种方法对 42份金黄色葡萄球菌阳性样品进行了肠毒素检测。结果RPLA法的金葡菌肠毒素检出率为 61.9% (P<0.05 )要高于ELFA法的检出率 50.0% (P <0.05 ) ,但检测时间长 ( 20h)。在实验中我们还发现 ,血浆凝固酶阴性的葡萄球菌也可产生肠毒素 ,其原因有待进一步的研究。