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.     

Preimplantation genetic diagnosis (PGD), a collaborative activity of clinical genetic departments and IVF centres

An Erratum has been published for this article in Prenatal Diagnosis 22 (5) 2002, 451. Preimplantation genetic diagnosis (PGD) requires the combined efforts of geneticists and workers in the field of reproductive medicine. This was studied on the basis of a questionnaire, sent to 35 members of the PGD Consortium of the European Society of Human Reproduction and Embryology (ESHRE). A reply was obtained from 20 centres. They represent the majority of activities in the field of PGD in the world. It is obvious that many of the activities (in vitro fertilisation, embryo culture and biopsy) take place in IVF units while others (counselling and diagnosis) are the responsibility of genetic diagnostic centres. The distances between both units vary considerably. In all but one centre sex determination is offered. Aneuploidy screening is offered in 13 out of 20 centres. PGD of translocations and other structural chromosome abnormalities is offered in all but one centre. The number of monogenic diseases offered varies considerably. In comparison to prenatal diagnosis PGD is more expensive. The majority of these costs are due to the IVF or ICSI procedure. The charges for PGD vary between about € 600 and € 4000. In 16 out of 20 centres the parents to be must sign an informed consent form. Copyright © 2001 John Wiley & Sons, Ltd.     

农村污水膜生物反应器系统中微生物群落解析

农村污水的随意排放和未加限制的灌溉回用可能会导致水源地环境污染加剧,并且其中潜在的病原微生物对周边淡水资源安全和农村居民的身心健康造成潜在的威胁.为了解农村污水的微生物群落组成,为后续污水灌溉的病原微生物风险评价提供理论依据,采用末端限制性片段长度多态性(T-RFLP)和16S r DNA基因克隆文库技术研究农村污水膜生物反应器(MBR)系统中的微生物群落多样性,并结合实时荧光定量PCR方法监测处理前后典型病原菌——弓形菌(Arcobacter spp.)以及总细菌数量的变化.从未处理的农村污水中获取的73个阳性克隆测序结果表明,其主要细菌类群为变形菌门(91.8%)、厚壁菌门(2.70%)、拟杆菌门(1.40%),以及部分不可培养细菌(4.10%).其中弓形菌属是ε-变形菌门的优势菌,也是农村污水中的优势菌株,占克隆总数的68.5%.TRFLP的结果表明,不同处理工艺阶段的主要细菌类群及丰度明显不同,调节池的物种丰度(S)、Shannon-Wiener(H)和物种均匀度(E)最高,分别为43.0、3.56和0.95.定量PCR结果显示未处理的农村污水中弓形菌数量高达(1.09±0.064 0)×1011copies·L-1,该结果与克隆文库结果均表明弓形菌在污水中确实占较高比例.与未处理的农村污水相比,处理后的出水中所监测的弓形菌及细菌总拷贝数分别减少了2~3个数量级,说明MBR处理系统在一定程度上能够去除微生物.处理后的再生水理化指标及指示菌卫生指标均符合农田灌溉用水标准,但其中残留病原微生物引发的健康风险仍需进一步评价.     

基于GOCI影像和水体光学分类的内陆湖泊叶绿素a浓度遥感估算

叶绿素a作为水质参数之一,常用来作为衡量水体富营养化程度的指示标准.利用从太湖及洞庭湖获取的326个实测数据,基于实测遥感反射率对水体光谱进行光学分类,结果表明所采集的样点可分为3种水体类型.结合GOCI的波段设置,建立了不同类型水体的叶绿素a浓度反演模型.水体类型一可以利用490 nm(3波段)和555 nm(4波段)来反演,水体类型二可利用660 nm(5波段)和443 nm(2波段),水体类型三利用745 nm(7波段)和680 nm(6波段).精度分析表明,分类后的平均相对误差明显下降,类型一为38.91%、类型二为24.19%、类型三为22.90%;类型一均方根误差为4.87μg·L-1、类型二为8.13μg·L-1、类型三为11.66μg·L-1;分类前后的总体平均相对误差由49.78%降低到29.59%,总体均方根误差由14.10μg·L-1降低到9.29μg·L-1,分类后反演精度得到了显著提高.利用2013年5月13日8景GOCI影像反演了太湖的叶绿素a浓度,结果表明,2013年5月13日太湖叶绿素a浓度日变化显著,高值区主要集中在竺山湾、梅梁湾、贡湖湾,低值区主要集中在湖心区以及南部区域,10:00以后太湖西南部沿岸的叶绿素a浓度显著降低.这种先分类后反演的方法对于二类水体的模型反演精度的提高具有重要作用.     

垃圾填埋场抗生素抗性基因初探

不同环境介质中抗生素抗性基因普遍存在,但是在垃圾填埋场中抗生素抗性基因尚无相关报道.本实验以西安江村沟垃圾填埋场为研究对象,采集不同方位不同深度垃圾样品,分析垃圾理化性质,用荧光定量PCR检测磺胺类抗生素抗性基因(sulⅠ和sulⅡ)、抗氯霉素类抗生素抗性基因(cat)、β-内酰胺类抗生素抗性基因(bla-SHV),以及四环素类抗生素抗性基因(tet W)等5种抗生素抗性基因的含量,以相关性分析垃圾理化性质与抗性基因的关联.结果表明,5种抗生素抗性基因均存在于垃圾中,基因拷贝数(以干土计)最大值分别为:(3.70±0.06)×108copies·g-1(sulⅡ)、(9.33±0.06)×106copies·g-1(sulⅠ)、(2.27±0.08)×105copies·g-1(tet W)、(3.68±0.09)×104copies·g-1(bla-SHV)和(1.39±0.10)×104copies·g-1(cat),说明垃圾填埋场是抗生素抗性基因潜在的储存库.抗性基因sulⅠ、sulⅡ和cat与含水率呈明显的正相关,同时sulⅠ和cat基因含量与p H值呈负相关.     

洪泽湖有毒和无毒微囊藻丰度及其与环境因子之间的相关分析

为了解洪泽湖有毒和无毒微囊藻丰度及其空间分布,于2014年8月对洪泽湖30个采样点水体的营养盐浓度和Calson富营养化指数(trophic state index,TSI)进行研究,同时采用实时荧光定量PCR技术测定了有毒和无毒微囊藻丰度.结果表明,洪泽湖水体的总氮和总磷浓度平均值分别为1.63 mg·L-1和0.11 mg·L-1,富营养化指数在58.1~73.6之间,洪泽湖水质呈富营养化状态;有毒微囊藻在洪泽湖广泛分布,其丰度在1.13×104~3.51×106copies·m L-1之间,总微囊藻丰度在1.06×105~1.10×107copies·m L-1之间,有毒微囊藻占总微囊藻的比例在8.5%~38.5%之间,平均值为23.6%,有毒微囊藻丰度及其比例均呈现出明显的空间差异性;相关分析结果显示,总微囊藻、有毒微囊藻和有毒微囊藻所占比例三者之间呈极显著正相关性(P0.01),总微囊藻和有毒微囊藻丰度与叶绿素a浓度和TSI有极显著正相关性(P0.01),与透明度有极显著负相关性(P0.01),有毒微囊藻所占比例与叶绿素a、总氮、总磷和TSI有极显著正相关性(P0.01),与TN/TP和透明度有极显著负相关(P0.01).因此,削减洪泽湖总氮、总磷浓度一方面可以降低水体富营养化水平,另一方面有利于抑制有毒微囊藻对无毒微囊藻的竞争优势.     

Comparison of two methods for detection of fecal indicator bacteria used in water quality monitoring of the Three Gorges Reservoir

Scientifically sound methods to rapidly measure fecal indicator bacteria are important to ensure safe water for drinking and recreational purposes.A total of 200 water samples obtained from the Three Gorges Reservoir during three successive one-year study periods(October 2009 to September 2012) were analyzed using multiple-tube fermentation(MTF)and most probable numbers combined with polymerase chain reaction(MPN–PCR).The MPN–PCR method was found to be significantly more sensitive than the MTF method for detecting Escherichia coli and Enterococcus spp.,and of equal sensitivity for detecting total coliforms when all surface water samples were grouped together.The two analytical methods had a strong,significant relationship,but MPN–PCR took only 12–18 hr,compared with the 3–8 days needed using the MTF method.Bacterial concentrations varied per sampling site but were significantly lower in the mainstream of the Yangtze River than those in the backwater areas of tributaries.The water quality of 85.8% of water samples from the mainstream was suitable for use as a centralized potable water source,while the water quality of 52.5% of water samples from the backwater areas was unsuitable for recreational activities.Relationships between fecal indicator bacteria showed significant correlation(r = 0.636–0.909,p 0.01,n = 200),while a weak but significant correlation was found between fecal indicators and water turbidity,water temperature,daily inflow,and total dissolved solids(r = 0.237–0.532,p 0.05,n = 200).The study indicated that MPN–PCR is a rapid and easily performed deoxyribonucleic acid(DNA)-based method for quantitative detection of viable total coliforms,E.coli,and Enterococcus spp.in surface water.