A practicable variant of the ion exchange method for the radiometric estimation of ornithine decarboxylase activity

Summary A known ornithine decarboxylase assay working with ion exchang separation of [³H]ornithine and [³H]putrescine has been revised. The assay can be performed in disposable 1.5 ml vessels with a total of four pipetting steps. The separation of enzyme substrate and product, respectively, requires 3 h per 50 samples. The detection limit is about 50 pmoles [³H]putrescine formed.     

In vitro systems in the study of peroxisomal protein import

Our level of understanding of peroxisome biogenesis in comparison with other cellular organelles is rudimentary, yet the fragments of information available indicate that the targeting and import of peroxisomal proteins occur by fundamentally different mechanisms. Genetic studies have identified a number of genes required for peroxisome assembly, but in most cases the functions of the gene products remain unknown. In vitro protein translocation systems have played a prominent role in unravelling the biochemistry of protein translocation into other organelles. This review considers some of the requirements for establishing a bona fide peroxisomal import assay and discusses the findings which have emerged as a result of using such experimental systems.     

Robust regression in biological assay: application to the evaluation of alternative experimental techniques

Summary Robust Huber type regression and testing of linear hypotheses are adapted to statistical analysis of parallel line and slope ratio assays. They are applied in the evaluation of results of several experiments carried out in order to compare and validate alternatives to animal experimentation based on embryo and cell cultures. Computational procedures necessary for the application of robust methods of analysis used the conversational statistical package ROBSYS. Special commands for the analysis of parallel line and slope ratio assays have been added to ROBSYS.     

应用PCR-RFLP法作脊灰炎病毒型内分析的研究

用PCR-RFLP法对近年来从麻痹病人分离到的26株脊灰炎病毒(PV)作了型内分析研究。16株PV₁的RFLP图式呈现多样性,毒株的图式可与Sabin1型疫苗株完全相同或不同,某些毒株亦可无电泳条带出现。7株PV₂与3株PV₃的RFLP图式则分别与疫苗株Sabin2型和3型完全相同。全面分析这些RFLP后,我们认为16株PV₁为野毒株,而7株PV₂及3株PV₃则可能是疫苗相似株。鉴于近年来广西的麻痹病例仍主要由PV₁所致。故此,加强突击普服OPV运动及作好分离毒株的型内分析已成为消灭脊灰炎的关键。     

对于核小体定位检测方法的比较研究

在真核细胞中,核小体是组成染色质的基本结构单位,是由DNA紧密缠绕在组蛋白八聚体上所形成的一个复合体结构。而DNA与组蛋白的结合并不是固定不变的,没有核小体结合的DNA区域易于各种调节蛋白的接近与结合。因此人们怀疑核小体的定位与基因的转录调节之间存在某种内在联系。对现行的核小体定位的检测方法进行了归类,并对其优缺点进行了分析整理。对更深入的探索核小体定位检测方法的应用有一定意义。     

胶体金免疫层析技术定量检测人绒毛膜促性腺激素

采用柠檬酸三钠还原法制备胶体金,用胶体金标记和双抗体夹心免疫层析方法,建立对人绒毛膜促性腺激素的快速定量检测方法．该检测方法可以在15min内完成对HCG的定量检测,灵敏度可达25mIU／mL,线性范围25～10000mIU／mL．与人促黄体生成素（hLH）、人促甲状腺激素（hTSH）、人卵泡刺激素均无交叉反应,所建立的检测人绒毛膜促性腺激素的胶体金免疫层析方法,能快速、灵敏、特异、准确地检测样品中的人绒毛膜促性腺激素,在异位妊娠的早期诊断及过程监控中具有较为广阔的应用前景．     

火试金技术的发展与应用

回顾了过去的60年,我国在火试金技术的发展与应用方面取得的成就,其中以铅试金法的发展与应用为主要内容及20世纪50年代以来为解决铂族元素的富集问题相继创立的有：铜铁镍试金法、锡试镍试金法、锡试金法、铜试金法、锑试金法和铋试金法等新的试金法进行综述。这些新试金法的建立大大丰富了火试金技术的内容,同时对贵金属的分析与贵金属地质标准物质的研制起到了极大的推动作用。本文拟就铅试金法、镍锍试金和锑试金法、铋试金法在地质样品中测定贵金属和在贵金属地质标准物质的研制工作中的应用与地位进行综述,并对锡试金法、铜铁镍试金法和铜试金法简要介绍。     

Rapid Screening for Insecticidal Compounds Using [3H] EBOB Binding Assay

Binding assay is a convenient, simple, inexpensive, and rapid method that can be used for rapid screening insecticidal compounds. The present study examines 6 compounds for their inhibition of [^3H] EBOB bound to housefly-head membranes, of which 3 compounds have potency for housefly-head membranes.     

斑点免疫结合法检测3种植物病毒

研究了改进的斑点免疫结合法在检测芜菁花叶病毒、烟草花叶病毒和小西葫芦黄化花叶病毒中的应用.无论是使用全抗血清还是纯化的免疫球蛋白,检测感染组织粗汁液中的病毒或纯化的病毒,均获得了满意的结果.同时进行的斑点免疫结合试验、酶联免疫吸附试验和免疫吸附电子显微术等的比较结果表明:无论是检测纯化病毒还是感染组织粗汁液中的病毒,斑点免疫结合法的检测敏感性均优于后2种方法,病毒的可测感度芜菁花叶病毒为0.65ng,烟草花叶病毒为0.39 ng,小西葫芦黄化花叶病毒为0.45 ng;应用碱性磷酸酶标记抗体及羟基吲哚磷酸盐和氮兰四唑为底物较为合适,这种底物可在室温下长期保存,并且反应产物为不褪色的紫色,易于观察和保存.     

酶标板上基于竞争杂交反应的几种miRNA序列的比色检测

多种miRNA序列的同时检测对癌症的预警与早期诊断具有重要的意义。本文基于生物素标记的miRNA和靶点miRNA与酶标板上固定的寡核苷酸探针之间的竞争反应,通过比色法来定量检测三种与胶质瘤相关的miRNA序列,即miR-182、miR-185和miR-381。通过亲和素与生物素之间的特异性相互作用,将亲和素标记的多聚辣根过氧化物酶(SA-polyHRP)衍生到微孔表面。通过检测HRP催化3,3′,5,5′-四甲基联苯胺和H2O2的显色反应来对三种靶点miRNA序列进行定量。方法最低检测浓度为10fmol/L,有望用于临床上miRNA的定量分析。碱基错配的结果表明该方法具有较好的选择性。