Digital PCR: a powerful new tool for noninvasive prenatal diagnosis?

Recent reports have indicated that digital PCR may be useful for the noninvasive detection of fetal aneuploidies by the analysis of cell-free DNA and RNA in maternal plasma or serum. In this review we provide an insight into the underlying technology and its previous application in the determination of the allelic frequencies of oncogenic alterations in cancer specimens. We also provide an indication of how this new technology may prove useful for the detection of fetal aneuploidies and single gene Mendelian disorders. Copyright © 2008 John Wiley & Sons, Ltd.     

高校事故非线性灰色预测模型及应用研究

通过调查和分析表明,近几年来湖南省高校事故中学生非正常死亡由自杀、交通、溺水等7项因素组成,且统计数据信息偏少。笔者将影响湖南省高校事故学生非正常死亡人数的各项因素作为一个系统,应用系统灰预测理论建立高校事故系统灰预测模型;将原始建模数据序列经过方根变换和数值初始化处理后,适合于建模的要求,构造GM(1,N)与GM(1,1)的嵌套模型;通过建模事中和事后的检验,预测湖南省2007年高校事故学生非正常死亡人数,预测值与实际值相差较小,模型精度较高;高校事故非线性灰色预测模型,为高校事故预测提供了一种应用方法并有一定的指导意义。     

应用荧光定量PCR技术监测玄武湖蓝藻水华

应用荧光定量PCR技术和显微计数法对玄武湖蓝藻水华进行了长期监测,结果表明,荧光定量PCR法可同步监测蓝藻、微囊藻和有毒微囊藻的数量,及时准确反映玄武湖蓝藻水华优势种群微囊藻和有毒微囊藻的动态变化。与显微计数法相比,具有需要的样品量少、时效性强、检出下限较低、自动化程度高等优势,可有效地应用于蓝藻水华的监测。     

The effect of thiacloprid formulation on DNA/chromosome damage and changes in GST activity in bovine peripheral lymphocytes

The potential genotoxic effect of thiacloprid formulation on bovine peripheral lymphocytes was evaluated using the comet assay and the cytogenetic endpoints: chromosome aberrations (CAs), sister chromatid exchanges (SCEs) and micronuclei (MNi). Whole blood cultures were treated with the insecticide at concentrations of 30, 60, 120, 240 and 480 μg mL^?1 for 24, 48 h and/or 2 h of incubation. A statistically significant increase in the frequency of DNA damage, as well as in unstable chromosome aberrations (% breaks) were found after exposure to the insecticide at concentrations ranging from 120 to 480 μg mL^?1 (P < 0.05, P < 0.01, P < 0.001). For the detection of stable structural chromosome aberrations (e.g., translocations) and numerical aberrations by the FISH method, three whole chromosome painting probes for bovine chromosomes 1, 5 and 7 (BTA1, BTA5 and BTA7) were used in our experiments. We observed numerical aberrations, but without any statistical significance. Regarding the sister chromatid exchanges, no significant elevation in the SCE frequencies was found after 24-h exposure to the insecticide. A dose-related response in the SCE induction was obtained in bovine cultures after the prolonged time of exposure (48 h) to thiacloprid formulation at concentrations ranging from 120 to 480 μg mL^?1 in each donor (P < 0.05, P < 0.01), which was associated with a reduction of the PI (P < 0.05, P < 0.01). The insecticide failed to produce MNi; however, a significant reduction of CBPI was observed. Using real-time PCR, a decrease in the expression of bovine glutathione S-transferase M3 (GSTM3) was detected at the lowest dose. The higher concentrations of thiacloprid formulation caused an increase in the mRNA expression.     

城市生活污水中志贺氏菌ipaH毒力基因的定量PCR检测

基于ipaH毒力基因的实时荧光定量PCR检测,建立适合城市污水中志贺氏菌的定量检测方法。使用从临床分离出的志贺氏菌构建重组质粒作为实时荧光定量PCR的标准品。在ipaH基因模板量2.58×100～2.58×106copy范围内具有良好的线性关系,每100 mL水样中含有2.58×101copy以上的ipaH基因即可被检出。从西安市生活污水分离得到2株野生型志贺氏菌,分析ipaH基因数量和菌体数量的关系,从而确定ipaH基因定量检测志贺氏菌的可行性。该方法灵敏、快速、特异性好,适用于城市生活污水中志贺氏菌的检测。     

一种新的检测土壤中壬基酚的荧光定量PCR方法

建立了一种新的外切酶保护-荧光定量PCR方法检测环境激素壬基酚,并将该方法应用于土壤样品中壬基酚的检测.壬基酚可以活化雌激素受体使之与包含特定序列的双链DNA结合,结合的DNA因受到蛋白质的保护可抵抗核酸外切酶Exo Ⅲ的消解而被保留,痕量的保留DNA可通过PCR扩增定量.根据此原理,首先从鱼肝脏中提取含雌激素受体的细胞溶质,与不同浓度的壬基酚结合使雌激素受体活化,形成配体-受体复合物;再设计合成特异性引物序列,采用常规PCR方法扩增制备双链结合DNA;使双链结合DNA与配体-受体复合物反应结合后,用核酸外切酶ExoⅢ和S_1核酸酶消解,去除未受到蛋白质保护的结合DNA.将消解后产物作为模板,进行荧光定量PCR扩增反应,从而建立C_t值与壬基酚质量浓度N(g/L)的标准曲线C_t=-1.828lgN+11.447.将该方法应用于土壤中壬基酚的检测,最低检测限达到2 pg/g,可用于检测大批量环境样品中壬基酚.     

荒漠绿洲土壤抗生素抗性基因分布特征及驱动机制

荒漠绿洲农田生态系统是干旱区环境下人类活动显著的复合生态系统.土壤微生物抗生素抗性与人类健康和生态平衡关系密切.研究荒漠绿洲环境不同土地利用类型模式下土壤抗生素抗性基因的多样性、分布特征和影响因素,对于评估干旱区土壤环境健康风险,促进绿洲农业生态的发展具有重要意义.采用高通量测序和高通量定量PCR技术对荒漠绿洲土壤微生物的群落结构和抗生素抗性基因多样性开展了研究,旨在探究干旱区土壤抗性基因的分布特征及其驱动机制.结果表明,从沙漠边缘到绿洲,荒漠沙生植物土壤、棉花地土壤、玉米地土壤、芦苇地土壤和湖泊沉积物中抗生素抗性的种类和丰度显著增加,与土地利用变化关系密切,农田土壤是抗性基因的重要存储库;荒漠绿洲土壤微生物群落与抗生素抗性基因显著相关,硫杆菌属(Thiobacillus)、沙漠细菌属(Pontibacter)、诺卡氏菌属(Nocardioides)、耐盐微杆菌属(Salinimicrobium)、土壤红杆菌属(Solirubrobacter)和链霉菌属(Streptomyces)等是各类抗性基因重要的潜在携带者;干旱区土壤中重(类)金属元素和可移动基因元件,与微生物群落共同塑造了抗生...