Prenatal diagnosis of autosomal dominant polycystic kidney disease using flanking DNA markers and the polymerase chain reaction

A prenatal diagnosis was carried out on a 9-week-old fetus at risk for autosomal dominant polycystic kidney disease (ADPKD). Ten members of the family were previously typed using five DNA markers linked to the PKD1 locus on chromosome 16, and one marker linked to the putative PKD2 locus on chromosome 2. The polymerase chain reaction (PCR) was used to amplify the D16S125 locus. Pairwise and multipoint lod scores indicated that the family was most likely segregating a PKD1 mutation. The fetus inherited the disease haplotype from the affected parent. Diagnostic accuracy was greater than 99 per cent, taking into account the possibility of genetic heterogeneity.     

Amplification of plasmid DNA bound on soil colloidal particles and clay minerals by the polymerase chain reaction

Polymerase chain reaction(PCR)was used to amplify a 600-base pair(bp)sequence of plasmid pGEX-2T DNA bound on soil colloidal particles from Brown soil(Alfisol)and Red soil(Ultisol),and three different minerals(goethite,kaolinite,montmorillonite). DNA bound on soil colloids,kaolinite,and montmorillonite was not amplified when the complexes were used directly but amplification occurred when the soil colloid or kaolinite-DNA complex was diluted,10- and 20-fold.The montmorillonite-DNA complex required at least 100-fold dilution before amplification could be detected.DNA bound on goethile was amplified irrespective of whether the complex was used directly,or diluted 10- and 20-fold.The amplification of mineral-bound plasmid DNA by PCR is,therefore,markedly influenced by the type and concentration of minerals used.This information is of fundamental importance to soil molecular microbial ecology with particular reference to monitoring the fate of genetically engineered microorganisms and their recombinant DNA in soil environments.     

Detection of Y chromosome-specific DNA in the plasma and urine of pregnant women using nested polymerase chain reaction

The present study was undertaken to evaluate a nested polymerase chain reaction (PCR) for detection of Y chromosome-specific fetal DNA in maternal plasma and urine of pregnant women during different gestational stages. DNA isolated from plasma and urine samples of 80 pregnant women (between 7 and 40 weeks' gestation) underwent amplification for Y chromosome-specific 198 bp DNA by nested PCR. The postpartum analysis of fetal gender showed that 55 women carried male and 25 female fetuses. Among the 55 women bearing male fetuses, Y chromosome-specific signals were detected in 53 (96%) plasma and 21 (38%) urine samples. Moreover, out of 25 women bearing female fetuses, 3 (12%) and 1 (4%) women had Y chromosome-specific signal in plasma and urine, respectively. Analysis of results with respect to gestational age revealed that there was no significant difference in the detection of Y chromosome-specific DNA between different trimesters in maternal plasma of women bearing male fetuses. These results showed that fetus-specific DNA was detected with high sensitivity (96%) and specificity (88%) in the maternal plasma by nested PCR, and therefore the method could be useful as a non-invasive procedure for fetal sex determination and prenatal diagnosis. Copyright © 2001 John Wiley & Sons, Ltd.     

A routine method for storing lymphocytes for repeated isolation of DNA

A protocol for easy storage and later expansion of lymphocyte populations is given. Compared with methods using transformed cell lines, the method has a number of advantages for repeated production of cells for the isolation of DNA in amounts sufficient for use in diagnostic DNA technology.     

富勒烯对PCR反应的抑制作用

以110bp单链DNA为模板,研究富勒烯(C60)对聚合酶链式反应(polymerase chain reaction,PCR)的影响.实验结果表明,随着C60浓度的增加,PCR反应被显著抑制;将Taq DNA聚合酶、单链DNA模板与C60孵育后,其PCR扩增产物均显著减少;增加PCR反应体系中的Taq DNA聚合酶量,可消除C60的抑制作用,但增加起始单链DNA模板的数量,效应不明显,上述研究结果说明,C60不仅可抑制Taq DNA聚合酶活性,同时对DNA模板也具有一定的损伤作用.     

DNA vaccines

Immunization by genes encoding immunogens, rather than with the immunogen itself, has opened up new possibilities for vaccine research and development and offers chances for new applications and indications for future vaccines. The underlying mechanisms of antigen processing, immune presentation and regulation of immune responses raise high expectations for new and more effective prophylactic or therapeutic vaccines, particularly for vaccines against chronic or persistent infectious diseases and tumors. Our current knowledge and experience of DNA vaccination is summarized and critically reviewed with particular attention to basic immunological mechanisms, the construction of plasmids, screening for protective immunogens to be encoded by these plasmids, modes of application, pharmacokinetics, safety and immunotoxicological aspects. DNA vaccines have the potential to accelerate the research phase of new vaccines and to improve the chances of success, since finding new immunogens with the desired properties is at least technically less demanding than for conventional vaccines. However, on the way to innovative vaccine products, several hurdles have to be overcome. The efficacy of DNA vaccines in humans appears to be much less than indicated by early studies in mice. Open questions remain concerning the persistence and distribution of inoculated plasmid DNA in vivo, its potential to express antigens inappropriately, or the potentially deleterious ability to insert genes into the host cell's genome. Furthermore, the possibility of inducing immunotolerance or autoimmune diseases also needs to be investigated more thoroughly, in order to arrive at a well-founded consensus, which justifies the widespread application of DNA vaccines in a healthy population.     

Prenatal diagnosis for dystrophia myotonica using the polymerase chain reaction

The polymerase chain reaction has been used to detect an abundant class of short repeat DNA families of the form (dC-dA)n.(dG-dT)n, known as microsatellites. These units are found throughout the human genome and have been characterized for several loci including APOC2 on chromosome 19ql2-ql3.2. The locus APOC2 is linked to the gene for dystrophia myotonica and a microsatellite within this locus was used to derive polymorphisms in a family to predict the inheritance of the disease. Chorionic villus sampling (CVS) was performed at 151/2 weeks' gestation. Following DNA extraction from the CVS material and parental blood samples, microsatellite analysis was carried out by the polymerase chain reaction.     

环境激素双酚A与DNA相互作用的电化学研究

采用电化学方法研究了双酚A与DNA间的相互作用。结果表明,DNA的存在使双酚A的氧化峰电流减小,且峰电位正移,双酚A与DNA的相互作用以嵌插作用为主。双链DNA（dsDNA）与双酚A的结合大于单链DNA（ssDNA）,结合比为1∶4,结合常数β为1.35×10^23。     

一种简便快速检测涕灭威对DNA损伤的方法

介绍一种简便、快速检测涕灭威对DNA损伤的方法,并将这种方法与其他几种检测DNA损伤的方法进行了比较.这种方法直接把菌体或组织匀浆液放入琼脂糖凝胶的加样孔穴中,在原位裂解后,在较高pH环境下进行电泳,根据DNA的量和损伤程度评价涕灭威对生物个体或生态系统产生的影响.     

乌鲁木齐大气PM2.5对质粒DNA的损伤研究

2012年1月~2012年12月采集乌鲁木齐大气PM2.5样品,使用质粒DNA评价法研究了不同季节PM2.5的氧化能力,并进行氧化性毒性与相应气象因素和质量浓度之间的相关性研究.结果表明,乌鲁木齐大气PM2.5的质量浓度具有冬季最高,春季和秋季次之,夏季最低的季节性变化特征;PM2.5全样和水溶部分氧化能力的季节差异较大,对质粒DNA的氧化性损伤具有冬季最大,春季和夏季之次,秋季最低.冬、春、夏、秋季大气PM2.5全样的TD30(PM2.5对质粒DNA造成破坏达到30%所需要的颗粒物的剂量)平均值分别为440,491,503,515μg/mL,水溶部分分别为474,721,666,600μg/mL.绝大部分PM2.5样品全样的TD30值均小于水溶部分样,表明全样的毒性大于相应的水溶部分样.全样TD30值与平均温度显著(P<0.05)正相关,表明寒冷的天气/季节可能造成PM2.5的高毒性.水溶样TD30值与风速显著(P<0.01)正相关,与相对湿度显著负相关.这表明,高的风速和低的相对湿度可能跟较低和较高的PM2.5的毒性有关.PM2.5氧化性损伤能力的大小与其质量浓度之间的相关性不明显,表明仅以颗粒物的质量浓度来评价大气颗粒物氧化性损伤能力大小的方法并不能真实地反映其对人体健康的危害程度,起决定作用的还是颗粒物的化学组成及其表面吸附的有害成分.     

一种分离纯化降解性质粒DNA的简便方法

控制微生物对某些环境污染物降解的质粒——降解性质粒,分子量较大,难于和染色体DNA分离。超螺旋质粒DNA在热变性后冷却即快速复性方面与染色体DNA不同。根据此性质,作者改进了McMaster的和Kado的方法,获得了良好效果。实验证明。经此法纯化的质粒具有转化活性,是研究化学污染物的降解性质粒的简便方法。     

聚合酶链式反应(PCR)用于检测环境水体指示菌的研究

对聚合酶链式反应（ｐｏｌｙｍｅｒａｓｅ ｃｈａｉｎ ｒｅａｃｔｉｏｎ，ＰＣＲ）快速检测水体指示菌－大肠菌群（ｔｏｔａｌ ｃｏｌｉｆｏｒｍ）和大肠杆菌（Ｅｓｃｈｅｒｉｃｈｉａ ｃｏｌｉ）进行了研究。采用的两对引物分别扩增了ＬａｃＺ基因的约２６０ｂｐ和ＵｄｉＡ基因的约１５０ｂｐＤＮＡ片段。引物Ⅰ和引物Ⅱ最低能分别扩增１０＾－６μｇ或１０＾－８μｇ基因组ＤＮＡ。整个检测工作在采集水样后５－６ｈ内完成。与     

一种高效提取焦化废水活性污泥总DNA的方法

对焦化废水活性污泥中微生物建立高质量的总DNA提取方法是开展分子生态学研究的重要前提.通过反复冻融-蛋白酶K-SDS、溶菌酶-反复冻融-SDS及溶菌酶-反复冻融-蛋白酶K-SDS这3种综合方法对焦化废水活性污泥总DNA进行提取,以OD260/OD280、OD260/OD230、产率、片段完整性、片段大小5个指标来评价样品总DNA的提取效果.结果表明,溶菌酶-反复冻融-蛋白酶K-SDS法所提取的总DNA的OD260/OD280值约为1.8,产率为1.90～16.30 μg·g-1,片断完整性好,主带清晰,大小约为23 kb,其PCR反应抑制物少,能够直接进行16S rRNA基因的PCR扩增.溶菌酶-反复冻融-蛋白酶K-SDS法能够为焦化废水活性污泥中微生物的分子生态学研究提供高质量的总DNA.     

Cell-free DNA screening in twin pregnancies: A more accurate and reliable screening tool

Objective

Outcome data from cell-free DNA (cfDNA) screening in twin gestations are limited. This study adds an appreciable number of confirmed outcomes to the literature, and assesses performance of cfDNA screening in twins over a 4.5-year period at one large clinical laboratory.

Method

Prenatal cytogenetic and SNP microarray results were cross-referenced with cfDNA results for twin pregnancies, yielding 422 matched cases. Using diagnostic results as truth, performance of cfDNA screening in this population was assessed.

Results

Of the 422 twin pregnancies with both cfDNA and diagnostic results, 3 specimens failed amniocyte analysis, and 48 samples (11.5%) were nonreportable from the initial cfDNA draw. Analysis of the 371 reportable samples demonstrated a collective sensitivity of 98.7% and specificity of 93.2% for trisomies 21/18/13. Positive predictive values (PPVs) in this study population, which is enriched for aneuploidy, were 78.7%, 84.6%, and 66.7% for trisomy 21, 18, and 13, respectively.

Conclusion

CfDNA screening in a cohort of twin pregnancies with matched diagnostic results showed superior performance compared to traditional serum biochemical screening in twins. This study adds to a growing body of evidence suggesting that cfDNA is an accurate and reliable screening tool for the major trisomies in twin pregnancies.