Detection of Plasmodium falciparum DNA in a microtitration plate-based hybridization test

A rapid DNA-test, depending on the affinity based hybrid collection principle, was developed for the detection of Plasmodium falciparum DNA from clinical specimens. In this method, hybridization takes place in solution and the hybrids are collected onto a solid phase for measurement. Two probes are used, one labelled with an affinity tag (biotin) and the other with a detectable label (32P). In the present test a single oligonucleotide complementary to a 21-base pair sequence which is highly repeated in the parasite genome served both as capture and detector probe. The test is a 2-h hybridization performed in streptavidin coated microtitration plate wells, onto which the labelled hybrids simultaneously bind. The sensitivity of the assay with a crude erythrocyte lysate specimen was 1.6 x 10(9) repeat units corresponding to about 160 parasites in one microliter blood. The results allowed quantification of the repeat sequences and thus estimation of the degree of parasitemia in clinical specimens.     

一种新型免疫层析技术在临床乳腺癌Her-2蛋白定量检测中的应用

建立一种靶点蛋白质快速定量检测方法。在原有侧向流动免疫层析技术的基础上,通过优化层析材料和纳米微球的均一性、改进检测区的检测方法,经逐点扫描技术,建立标准浓度曲线,以达到对临床靶点蛋白质的定量检测。以乳腺癌组织中的Her2表达为例,通过对已知浓度样品的检测,验证本技术方法的准确度大于96%。另外,以蛋白质免疫印迹作为组织中特定蛋白质检测金标准,分析临床肿瘤组织中Her2蛋白的含量,其准确率也达到95.5%,而免疫组织化学方法检测准确率仅为69.58%。新型免疫层析法检测结果与靶向治疗患者的愈后密切相关(P<0.01)。改进后的新型免疫层析方法能够准确地对临床靶点蛋白质进行定量检测,而且结合侧向流动技术的简单、快速和易用性,这种新型检测方法可以广泛应用于临床组织标本、血液标本和体液标本中靶点蛋白质的临场定量检测,在一定程度上可以替代免疫组化技术。     

Functional Heterologous Production of Reductive Dehalogenases from Desulfitobacterium hafniense Strains

The anaerobic dehalogenation of organohalides is catalyzed by the reductive dehalogenase (RdhA) enzymes produced in phylogenetically diverse bacteria. These enzymes contain a cobamide cofactor at the active site and two iron-sulfur clusters. In this study, the tetrachloroethene (PCE) reductive dehalogenase (PceA) of the Gram-positive Desulfitobacterium hafniense strain Y51 was produced in a catalytically active form in the nondechlorinating, cobamide-producing bacterium Shimwellia blattae (ATCC 33430), a Gram-negative gammaproteobacterium. The formation of recombinant catalytically active PceA enzyme was significantly enhanced when its dedicated PceT chaperone was coproduced and when 5,6-dimethylbenzimidazole and hydroxocobalamin were added to the S. blattae cultures. The experiments were extended to D. hafniense DCB-2, a reductively dehalogenating bacterium harboring multiple rdhA genes. To elucidate the substrate spectrum of the rdhA3 gene product of this organism, the recombinant enzyme was tested for the conversion of different dichlorophenols (DCP) in crude extracts of an RdhA3-producing S. blattae strain. 3,5-DCP, 2,3-DCP, and 2,4-DCP, but not 2,6-DCP and 3,4-DCP, were reductively dechlorinated by the recombinant RdhA3. In addition, this enzyme dechlorinated PCE to trichloroethene at low rates.     

独岛枝芽胞杆菌MCCC 1A00493产生的4-乙烯基苯酚鉴定和作用南方根结线虫特性研究

【目的】根结线虫危害严重,难防难控,前期研究发现一株深海来源的独岛枝芽胞杆菌(Virgibacillus dokdonensis) MCCC 1A00493对南方根结线虫具有良好的体外拮抗效果,本研究分离鉴定菌株发酵液中杀线虫活性物质,对其作用线虫的多种模式进行研究,为菌株有效控制植物病原线虫的应用奠定理论基础。【方法】采用硅胶柱层析和半制备高效液相色谱对独岛枝芽胞杆菌发酵上清液中的杀线虫活性物质进行分离纯化,采用液相色谱质谱联用、核磁共振对纯化物进行结构鉴定;并对其趋避、诱杀、熏杀和卵孵化抑制活性进行检测。【结果】结构鉴定确定独岛枝芽胞杆菌MCCC 1A00493发酵液中分离的杀线虫活性物质为4-乙烯基苯酚。4-乙烯基苯酚对线虫具有触杀、熏杀、卵孵化抑制和高浓度驱避低浓度引诱活性,15μg/mL 4-乙烯基苯酚72 h触杀线虫校正死亡率为71.23%±9.06%;20 mg/mL 4-乙烯基苯酚24 h熏杀线虫死亡率达100%;100μg/mL4-乙烯基苯酚作用线虫卵10d后,卵孵化抑制率达66.2%;在琼脂平板上,10mg/mL的4-乙烯基苯酚对线虫有驱避作用,1 mg/mL的4-...     

荒漠灌丛土壤动物分布及其生态功能

在干旱、半干旱荒漠生态系统中,灌丛作为一种重要的植被类型,其独特的形态和生理适应特性能够有效促进退化生态系统结构与功能的恢复。土壤动物是荒漠生态系统中不可或缺的重要组成部分,对促进灌丛"肥岛"演变具有重要的生态作用,有利于灌丛生态功能的发挥及退化生态系统的恢复。近年来,国内外学者对荒漠灌丛微生境土壤动物的研究逐步深入,取得大量的研究成果。在此基础上,首先综述荒漠灌丛微生境土壤动物群落分布和生态功能,总结灌丛与土壤动物分布间作用关系的数学模型,针对荒漠灌丛土壤动物研究中存在的问题提出了未来可能的研究方向和建议。