The Escherichia coli O157:H7 DNA detection on a gold nanoparticle-enhanced piezoelectric biosensor

This paper presents development of a quartz crystal microbalance (QCM) biosensor for real-time de- tection of E. coli O157:H7 DNA based on nanogold particles amplification. Many inner Au nanoparticles were immobilized onto the thioled surface of the Au electrode, then more specific thiolated sin- gle-stranded DNA (ssDNA) probes could be fixed through Au-SH bonding. The hybridization was in- duced by exposing the ssDNA probe to the complementary target DNA of E. coli O157:H7 gene eaeA, then resulted in a mass change and corresponding frequency shifts ( △f ) of the QCM. The outer avidin-coated Au nanoparticles could combine with the target DNA to increase the mass. The electro- chemical techniques, cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were adopted to manifest and character each step. The target DNA corresponding to 2.0×103 colony forming unit (CFU)/mL E. coli O157:H7 cells can be detected by this biosensor, so it is practical to develop a sensitive and effective QCM biosensor for pathogenic bacteria detection based on specific DNA analy- sis. The piezoelectric biosensing system has potential for further applications, such as food safety and environment monitoring, and this approach lays the groundwork for incorporating the method into an integrated system for in-field bacteria detection.     

大肠杆菌O157:H7基因组研究进展

对大肠杆菌O157：H7基因组的结构、特点等近年来的研究进展进行了综述。     

肠出血性大肠杆菌O157:H7感染小鼠动物模型的初步建立

目的　建立肠出血性大肠杆菌O15 7:H7感染小鼠动物模型。方法　选用离乳小鼠随机分组 ,先以丝裂霉素和萘啶酮酸处理 ,提高小鼠对肠出血性大肠杆菌O15 7的敏感性 ,再分别以不同剂量口服接种肠出血性大肠杆菌O15 7:H7EDL933W ,进行临床和病理学检查。结果　利用对O15 7不易感的小鼠 ,复制出人类感染O15 7所出现的主要病理变化和部分临床症状 ,初步建立了动物模型。讨论　该动物模型对探索肠出血性大肠杆菌O15 7:H7的感染机制、毒力因子的作用有重要意义 ,并为O15 7:H7的疫苗侯选株安全性的评价打下初步基础。     

大肠杆菌O157∶H7胶体金免疫层析快速检测法的建立

建立一种大肠杆菌O157∶H7的胶体金免疫层析快速筛查方法.利用胶体金免疫层析技术,采用双抗体夹心法检测大肠杆菌O157∶H7,对该法进行敏感性、特异性和食品样品的适用性分析.该法能在10 min内完成检测;该试纸条仅与O157∶H7阳性样品发生特异性反应;大肠杆菌O157∶H7的最低检出浓度为1×105 cfu/mL.新建立的大肠杆菌O157∶H7胶体金免疫层析试验简便、快速,特异性和灵敏度较好,适用于现场样品的快速筛查.     

冻融循环下大肠杆菌O157:H7在长春市南湖岸边水中的存活行为研究

北方地区的初春、晚秋,水体表层的夜冻昼融过程是典型特征。冻融循环过程影响水体自身理化性质,理化性质的变化对水中微生物,尤其是潜在的致病微生物的存活也会产生一定的影响。实验在室内模拟了大肠杆菌O157:H7在冻融循环条件下,在长春南湖水中的动态存活特征;并通过线性模型和Weibull模型,计算大肠杆菌O157:H7在污染水体后衰减达到最低检测限的时间(ttd);并运用多元线性回归进行相关性分析,找出影响实验菌种存活的关键因素。研究表明,南湖水的p H和氨氮与大肠杆菌O157:H7的存活相关。研究结果可为调查大肠杆菌O157:H7在湖水中的存活趋势,评估其可能带来的环境健康风险,为制定相应的环境管控措施提供科学依据。     

肠出血性大肠埃希菌O157:H7紧密黏附素C300与LTB融合蛋白B细胞表位预测

目的 对肠出血性大肠埃希菌O157:H7紧密黏附素C300与黏膜佐剂不耐热肠毒素(LTB)融合蛋白二级结构及B细胞识别表位进行预测,为开发新型疫苗提供理论依据.方法 采用DNA star软件中的Protean预测软件对Intimin C300与LTB融合蛋白的亲水性指数、β-转角、柔韧性、表面可及性和抗原指数进行预测....     

Analysis of components of conserved “backbone sequences” among genomes of<Emphasis Type="Italic">Shigella</Emphasis> spp. strains

Difference in the genomic compositions of prokaryotes is the basis of the diversity in their biological characters. However, besides their flora- or strain-specific genes, those floras with closer relationship in the evolution also have conserved “backbone sequences”, which reveal the marks of their origin and evolution, and these “backbone sequences” are just the basis of their elementary living abilities and common biological properties. Shigella is very closely related to E. coli in the origin and evolution, and may turn out to belong to the same genus. In this study, a microarray containing E. coli K-12 whole genome and SG01 specific ORFs is used to investigate the genomic components of four Shigella strains. The results indicate that 16%–22% K-12 ORFs sequences are not detected in the genome of Shigella strains while the genome of Shigella contains at least 2800 conserved ORFs, which compose the common “backbone sequences”. Advanced analysis indicated that the “backbone sequences” are the essential components in maintaining the normal physiological activities of intestinal bacteria. Furthermore, only 20% SGO1-specific ORFs exist in other strains simultaneously, which demonstrate the great genome heterogeneity and the genetic diversity among the strains. the first two authours made equal contribution to this work.     

快速检验食品中沙门氏菌和O157:H7大肠杆菌

利用沙门氏菌快速检验装置(Oxoid公司出口）和可视免疫沉淀物（VIP）试剂盒（BioControlSystems公司出口）对广西桂林市的七星区某农贸市场的20种食品进行了沙门氏菌和O157：H7大肠杆菌的快速抽样检验,结果发现,其中的11种食品的沙门氏菌呈阳性反应,4种食品的O157：H7大肠杆菌也呈阳性反应,与目前国内普遍使用的传统检验方法相比,该方法具有快速、简便、高效、准确的特点。     

基于金纳米棒/多壁碳纳米管-壳聚糖复合膜电极的肌红蛋白直接电化学和电催化研究

采用金纳米棒(AuNRs)/多壁碳纳米管-壳聚糖(MWCNTs-Chit)复合膜促进肌红蛋白在电极上的直接电子转移,并用于构建H2O2生物传感器.首先将金纳米棒固定到玻碳电极表面,然后把MWCNTs-Chit分散溶液和肌红蛋白(Mb)固载到玻碳电极上,得到MWCNTs-Chit/Mb/AuNRs复合膜电极.通过循环伏安法对膜电极进行表征,在pH=7.0磷酸缓冲溶液中,Mb表现出一对峰形良好且可逆的氧化还原峰,其中氧化峰和还原峰电位分别为-0.291 V、-0.235 V,式电位(Eθ’)为-0.263 V.与此同时还探讨了修饰电极的电催化活性,结果表明其对H2O2具有良好的电催化还原作用,可作为检测H2O2的生物传感器.传感器对H2O2的米氏常数为0.0494 mM,线性范围为5.0×10-5～5.0×10-3M(R=0.986 7,n=10),检测限为3.2×10-6M(信噪比为3).     

2种短周期人工林的林下植物多样性与生物量研究

2002年3月在经过炼山同一坡面相同海拔高度的杉木人工林采伐迹地上营造尾巨桉(Eucalyptus urophylla×E.grandis)和厚荚相思(Acacia crassicarpa)人工林.2007年10～12月以灌草坡自然恢复植被作为对照,设置样方调查2种短周期人工林的林下植物多样性和生物量.结果表明,2种短周期人工林的林下植物物种均为27种,均比对照的43种减少37.2%;对于灌木层,尾巨桉和厚荚相思人工林的物种丰富度指数分别比对照的减少36.4%和18.2%,Shannon-Wiener指数分别比对照的增加8.9%和27.4%,Pielou均匀度指数分别比对照增加27.5%和36.3%;对于草木层,尾巨桉和厚荚相思人工林物种丰富度指数分别比对照的减少38.1%和57.1%,尾巨桉Shannon-Wiener指数比对照的减少72.7%、厚荚相思比对照的增加29.8%,尾巨桉人工林Pielou均匀度指数比对照的减少67.6%,而厚荚相思人工林比对照的增加79.9%;尾巨桉和厚荚相思人工林的林下植物生物量分别比对照的减少40.8%和83.2%,2种短周期人工林的林下灌草植物生物量分配分别与对照的分配规律相一致.     

麻栗坡半蒴苣苔：越南苦苣苔科植物国家级分布新记录

麻栗坡半蒴苣苔Hemiboea malipoensis Y.H. Tan是最近才得以被描述的、原产自中国云南麻栗坡县的苦苣苔科新种,最近在越南北部河江省(Ha Giang Province)的全坝区(Quan Ba District)也发现了该种。该新记录种的凭证标本保存在越南生态与生物资源研究所标本馆(HN)和俄罗斯科马罗夫植物研究所(LE)。本文亦同时提供了本种的详细形态描述、彩色图片、物候、生态学、保育现状等信息和目前越南已知的半蒴苣苔属植物的检索表。     

产丁醇大肠杆菌工程菌的构建

【目的】为了在大肠杆菌(Escherichia coli)中导入改良的丁醇合成途径,使非生产菌株大肠杆菌具备产丁醇的能力。【方法】克隆大肠杆菌乙酰转移酶基因atoB和丙酮丁醇梭菌(Clostridium acetobutylicum)丁醇合成途径关键酶基因(crt、hbd、adhE),构建多顺反子表达质粒pSE380-atoB-adhE-crt-hbd;克隆齿垢密螺旋体(Treponema denticola)反式烯酰辅酶A还原酶基因ter,构建表达质粒pSTV29-ter,并将双质粒导入到大肠杆菌。【结果】构建的工程菌能半厌氧发酵产微量丁醇,产量为0.08g/L。【结论】大肠杆菌中的丁醇合成途径导入成功,构建了产丁醇的大肠杆菌工程菌。     

Microcalorimetric study on the antibiotic activity of clarithromycin and erythromycin

By using LKB-2277 Bioactivity Monitoring System, the heat effect changes in the process of inhibitory action of clarithromycin and erythromycin onEscherichia coli at 37°C were determined. Quantitative analysis showed that relationship between antibiotic concentrationc and rate contantk ofEscherichia coli growth, and half inhibitory ratio concentration IC₅₀: clarithromycin:k=0. 030 03–1. 1736×10⁻³ c, 8. 45 mg ·L⁻¹; erythromycin:k=0.031 08–8.4657×10⁻⁴ c, 14. 45 mg·L⁻¹. As a result of the microcalorimetry experiments, it not only indicated that antibacterial activity of clarithromycin was stronger than that of erythromycin, but also reported the changeable features of thermodynamics of the bacterial cell in biological, biochemical and metabolic process under different drug action. Foundation item: Supported by Natinal Natural Science Fundation of China (29973030), Natural Science Fundation of Hubei Province (98J052) and Post-doctoral Science Fundation of China Biography, SHEN Xue-song (1956-), Associate professor Research direction: biothermochemistry.     

巨尾桉枝叶水浸提液对3种作物种子萌发的影响

为了解巨尾桉（Ｅｕｃａｌｙｐｔｕｓｇｒａｎｄｉｓ＊Ｅ．ｕｒｏｐｈｙｌｌａ）内含物对其他植物的他感作用,分别用巨尾桉的新鲜枝叶和枯花叶的水浸认进行水稻、蕹菜（Ｉｐｏｍｏｅａａｑｕａｔｉｃａ）和菜苔（ＢｒａｓｓｉｃａｐａｒａｃｈｉｎｅｎｓｉｓＢａｉｌｅｙ）花种子发芽试验,经鲜液供水的水稻种子平均发芽率、发芽势分别为９５％、６０％略低于对照的９７％和６２％;蕹菜种子平均发芽率、发芽势为９３％、６２％,略     

Sensing Escherichia coli O157：H7 via frequency shift through a self-assembled monolayer based QCM immunosensor

By means of the specific immuno-recognition and ultra-sensitive mass detection, a quartz crystal microbalance （QCM） biosensor for Escherichia coli O157：H7 detection was developed in this work. As a suitable surfactant, 16-mercaptohexadecanoic acid （MHDA） was introduced onto the Au surface of QCM, and then self-assembled with N-hydroxysuccinimide （NHS） raster as a reactive intermediate to provide an active interface for the specific antibody immobilization. The binding of target bacteria with the immobilized antibodies decreased the sensor＇s resonant frequency, and the frequency shift was correlated to the bacterial concentration. The stepwise assembly of the immunosensor was characterized by means of the electrochemical techniques. Using the immersion-dry-immersion procedure, this QCM biosensor could detect 2.0×10^2 colony forming units （CFU）/ml E. coli O157：H7. In order to reduce the fabrication time, a polyelectrolyte layer-by-layer self-assembly （LBL-SA） method was adopted for fast construction. Finally, the reproducibility of this biosensor was discussed.     

多环芳烃K区和L区理论指数的计算

用K区和L区理论指数预测多环芳烃的致癌活性时,要计算分子的碳定域能(C、L、E)_(min),邻位定域能(O、L、E)和对位定域能(P、L、E)。本文以苯分子为例,对计算方法作了具体介绍。     

Plasmonic biosensors and nanoprobes based on gold nanoshells

Gold nanoshells (GNSs), consisting of a dielectric core coated with gold, have gained extensive attention as they show readily tunable optical properties and good biocompatibility. As highly sensitive and label-free optical biosensors with wide applications, GNSs have been investigated in many fields including drug delivery, immunoassay, cancer treatment, biological sensing and imaging. Taking advantage of the adjustability of the local surface plasmon resonance (LSPR) and the sensitivity of the surfaceenhanced Raman scattering (SERS) signal of GNSs, we have developed diverse applications including plasmonic biosensors and nanoprobes based on GNSs. In this review we introduce plasmonic and electromagnetic properties and fabrication methods of GNSs. We describe research progress in recent years, and highlight several applications of GNSs developed by our group. Finally we provide a brief assessment of future development of GNSs as plasmonic materials that can be integrated with complementary analytical techniques.     

烟碱降解菌DAB2菌株的筛选、鉴定和降解特性

以烟碱为唯一碳源,从湖北省襄阳烟叶种植土壤中分离得到了一株烟碱降解菌,命名为DAB2,经形态观察、生理生化分析和16S rDNA序列分析,初步鉴定该菌株为中间苍白杆菌(O.intermedium).当培养基中烟碱质量浓度为1.0 g/L,DAB2菌株生长良好,烟碱降解率为88.42%;当烟碱质量浓度为3.0 g/L,降解率为52.46%;当烟碱质量浓度高于6.0 g/L时,菌株生长和烟碱降解率均呈下降趋势,烟碱降解率低于10.36%.DAB2菌株在烟碱质量浓度为1.0 g/L的烟碱培养基中培养,不同时间样品紫外扫描显示:0 h只有259nm处有吸收波峰,8 h时259nm处波峰降低,在230nm和290nm处出现了新峰,表明产生了中间产物.