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实时荧光定量PCR 2

ATP荧光检测 1

COVID-19 1

DNA计算 1

PCR核酸检测 1

cDNA克隆 1

促黄体激素（LH） 1

分子诊断 1

半滑舌鳎 1

图顶点着色问题 1

微反应器 1

抗原检测 1

控制 1

放射免疫测定 1

检测策略 1

测量 1

生物检测 1

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662 A/G gene variation in human tumor necrosis factor receptor superfamily, member 9 (TNFRSF9)

QU Yanchun, YANG Ze, SUN Liang, JI Linong

《医学前沿（英文）》 2008年 第2卷 第3期   页码 283-285 doi: 10.1007/s11684-008-0053-7

摘要： The aim of this paper is to report a new coding variance of the gene, a candidate for autoimmune diseases. We found the variation in two families with type 2 diabetes mellitus by D-HPLC mutation screening method and confirmed our results by direct sequencing and PCR-RFLP. Although without changing the amino acid coding, the variance may have an effect on codon usage and play a role in disease development, such as type 2 diabetes mellitus. However, we cannot define the role of this variance because the frequency of the minor allele is low in the Chinese population and no homozygote of the variance was found. More research in multiple populations will be necessary to define the role of this variance.

关键词： D-HPLC mutation     development     autoimmune     PCR-RFLP     candidate    

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Spatial and seasonal variations in bacterial communities of the Yellow Sea by T-RFLP analysis

Hongyuan WANG, Xiaolu JIANG, Ya HE, Huashi GUAN

《环境科学与工程前沿（英文）》 2009年 第3卷 第2期   页码 194-199 doi: 10.1007/s11783-009-0018-3

摘要： Four typical coastal sites (rocky shore, sandy shore, mud flat shore, and artificial harbor) at the Yellow Sea were chosen to investigate the spatial and seasonal variations in bacterial communities. This was accomplished by using terminal restriction fragment length polymorphism (T-RFLP) analysis of PCR amplified 16S rDNA fragments. Two kinds of tetrameric restriction enzymes, I and I, were used in the experiment to depict the bacterial community diversity in different marine environments. It was found that the community compositions digested by the two enzymes separately were different. However, the results of bacterial community diversity derived from them were similar. The MDA analysis results of T-RFLP profiles coming from I and I both exhibited a significant seasonal community shift for bacteria and a relatively low spatial variation among the four locations. With I as the sample, the pair wise -tests also revealed that variations were minor between each pair of marine environments, with ranging from 0.198 to 0.349. However, the bacterial community structure in the mud flat site depicted a larger difference than each of the other three sites ( ranging from 0.282 to 0.349).

关键词： terminal restriction fragment length polymorphism (T-RFLP)     bacterial community structure     marine microbial diversity     HhaI     MspI    

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Using pyrosequencing and quantitative PCR to analyze microbial communities

Husen ZHANG

《环境科学与工程前沿（英文）》 2011年 第5卷 第1期   页码 21-27 doi: 10.1007/s11783-011-0303-9

摘要： New high-throughput technologies continue to emerge for studying complex microbial communities. In particular, massively parallel pyrosequencing enables very high numbers of sequences, providing a more complete view of community structures and a more accurate inference of the functions than has been possible just a few years ago. In parallel, quantitative real-time polymerase chain reaction (QPCR) allows quantitative monitoring of specific community members over time, space, or different environmental conditions. In this review, the principles of these two methods and their complementary applications in studying microbial ecology in bioenvironmental systems are discussed. The parallel sequencing of amplicon libraries and using barcodes to differentiate multiple samples in a pyrosequencing run are explained. The best procedures and chemistries for QPCR amplifications are also described and advantages of applying automation to increase accuracy are addressed. Three examples in which pyrosequencing and QPCR were used together to define and quantify members of microbial communities are provided: in the human large intestine, in a methanogenic digester whose sludge was made more bioavailable by a high-voltage pretreatment, and on the biofilm anode of a microbial electrolytic cell. The key findings in these systems and how both methods were used in concert to achieve those findings are highlighted.

关键词： polymerase chain reaction (PCR)     microbial communities     pyrosequencing     gut     microbial fuel cell     sludge    

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低成本PCR微反应器阵列的温度测量与控制技术

李现明,张玉林,李建田

《中国工程科学》 2007年 第9卷 第7期   页码 76-81

摘要：

设计了一种用于聚合酶链式反应的低成本微反应器，其内部无微加热器、微传感器。针对这种低成本的微反应器，提出了一种温度测量与控制方案，其特点是将各反应器温度的集中控制与分散控制有机结合，能够同时完成上百只微反应器快速、准确的温度循环。重点阐述了温度传感器负荷效应的提取与补偿、动态误差补偿、微反应器温度间接测量误差评定、微反应器温度串级控制策略。

关键词： 微反应器     测量     控制     聚合酶链式反应    

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Erratum to: Using pyrosequencing and quantitative PCR to analyze microbial communities

Husen ZHANG

《环境科学与工程前沿（英文）》 2011年 第5卷 第3期   页码 488-488 doi: 10.1007/s11783-011-0342-2

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Safety evaluation of microbial pesticide (HaNPV) based on PCR method

Miao Zhao, Shufei Li, Qinghong Zhou, Dianming Zhou, Ning He, Zhiyong Qian

《化学科学与工程前沿（英文）》 2019年 第13卷 第2期   页码 377-384 doi: 10.1007/s11705-018-1777-9

摘要： Microbial pesticides can prevent and control diseases and pests of crops, and has become one of the important measures to ensure food and environmental safety. However, the potential harm of microbial pesticides to humans and animals is a serious concern at home and abroad. In this paper, we have investigated the infectivity and pathogenicity of a representative of viral microbial pesticides, helicoverpa armigera nuclear polyhedrosis virus (HaNPV), by specific and highly sensitive polymerase chain reaction technology. The results show that HaNPV can be gradually cleared in a short time after getting into blood of experimental rats, and does not infect other tissues or organs of animals; also indicate that the test subjects are not infectious to experimental rats after intravenous injection of HaNPV. Our method has good specificity and repeatability, and could provide an important reference for establishment of infectivity and pathogenicity detection methods for viral microbial pesticides in future.

关键词： microbial pesticides     HaNPV     acute intravenous injection     infectivity     pathogenicity    

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Current molecular biologic techniques for characterizing environmental microbial community

Dawen GAO, Yu TAO

《环境科学与工程前沿（英文）》 2012年 第6卷 第1期   页码 82-97 doi: 10.1007/s11783-011-0306-6

摘要： Microbes are vital to the earth because of their enormous numbers and instinct function maintaining the natural balance. Since the microbiology was applied in environmental science and engineering more than a century ago, researchers desire for more and more information concerning the microbial spatio-temporal variations in almost every fields from contaminated soil to wastewater treatment plant (WWTP). For the past 30 years, molecular biologic techniques explored for environmental microbial community (EMC) have spanned a broad range of approaches to facilitate the researches with the assistance of computer science: faster, more accurate and more sensitive. In this feature article, we outlined several current and emerging molecular biologic techniques applied in detection of EMC, and presented and assessed in detail the application of three promising tools.

关键词： molecular biological technique     microbial community     denaturing gradient gel electrophoresis (DGGE)     terminal restriction fragment length polymorphism (T-RFLP)     environmental applications    

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A pooling strategy of a PCR-based assay to detect Angiostrongylus cantonensis in snail intermediate host

Fu-Rong WEI MS, Shan LV PhD, He-Xiang LIU, Ling HU, Yi ZHANG MS,

《医学前沿（英文）》 2010年 第4卷 第2期   页码 204-207 doi: 10.1007/s11684-010-0043-4

摘要： Pooling field specimens could reduce the number of assay and thus increase the efficiency in detecting and screening pathogen infections by polymerase chain reaction (PCR)-based assay. We investigated a pooling strategy in diagnosis of in Two settings of specimens were prepared, divided into portions and detected by multiplex PCR. Specimens A was 0.4490 g positive lung tissue of 28 larval nodes from four snails mixed with 1.310€g negative lung tissue from six snails and divided into 32 portions. Specimens B was 0.5448 g positive lung tissue with 26 larval nodes from two snails mixed with 1.092 g negative lung tissue from seven snails and divided into 48 portions. Repeated sampling was performed and sample size-accumulated positive rate curves were drawn. According to the sample size-accumulated positive rate curves, the appropriate sample size of the two specimens were 18 and 15, respectively, which is 0.36–0.58 to the total sample size. These test characteristics and the relevant factors to the sample size would need to be determined in much larger studies and more appropriately in field populations. The result indicates that the number of larval node is not the most important, nor the only factor to the sample size. It also implies the feasibility to detect in by pooling strategies.

关键词： Angiostrongylus cantonensis     Pomacea canaliculata     multiplex polymerase chain reaction     diagnosis     pooling    

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生物检测及分子诊断高端装备的研制及应用

彭年才,李 磊,李 政,苗保刚,李 明,赵 垚,蒋庄德

《中国工程科学》 2013年 第15卷 第1期   页码 57-62

摘要：

以生物检测与分子诊断技术发展为背景，总结了团队在医学诊断、疾控、食品安全、检疫检验、生物学医学科学研究等领域的分子诊断高端装备的研制情况，重点介绍了实时荧光定量聚合酶链式反应（PCR）分子诊断仪关键技术的研发及仪器工程化技术

关键词： 生物检测     分子诊断     实时荧光定量PCR     ATP荧光检测    

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基于抗原和聚合酶链式反应的组合策略对模拟社区内遏制COVID-19传播效果的定量分析 Article

黄蔷如, 孙艳侠, 贾萌萌, 张婷, 陈方圆, 姜明月, 王晴, 冯录召, 杨维中

《工程（英文）》 2023年 第28卷 第9期   页码 234-242 doi: 10.1016/j.eng.2023.01.004

摘要： 研究结果表明抗原检测及其组合策略比聚合酶链式反应（polymerase chain reaction, PCR）相关策略更高效，更有助于降低医院床位和重症监护病房床位需求。较短的结果返回时间是抗原检测策略的主要优势所在，也是优化PCR策略的关键因素。优化调整检测结果返回时间后，低频次的检测策略的效果可以媲美每日连续检测策略的效果。

关键词： COVID-19     抗原检测     PCR核酸检测     检测策略    

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半滑舌鳎促黄体激素基因克隆和表达分析及其血清浓度测定

柳学周,史宝,王珊珊,徐永江,李晓晓

《中国工程科学》 2014年 第16卷 第9期   页码 50-60

摘要： 实时荧光定量组织表达分析表明，LH mRNA在脑、垂体、卵巢等组织中表达，垂体中表达量最丰富，其他组织尤以脑、性腺表达量较高，推测半滑舌鳎LH可能具有广泛的垂体外生理功能；实时荧光定量PCR方法对繁殖周期雌性半滑舌鳎

关键词： 半滑舌鳎     促黄体激素（LH）     cDNA克隆     实时荧光定量PCR     放射免疫测定    

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Community dynamics of ammonia oxidizing bacteria in a full-scale wastewater treatment system with nitrification stability

Xiaohui WANG, Xianghua WEN, Hengjing YAN, Kun DING, Man HU

《环境科学与工程前沿（英文）》 2011年 第5卷 第1期   页码 92-98 doi: 10.1007/s11783-010-0254-6

摘要： To determine whether the functional stability of nitrification was correlated to a stable community structure of ammonia oxidizing bacteria (AOB) in a full-scale wastewater treatment plant, the AOB community dynamics in a wastewater treatment system was monitored over one year. The community dynamics were investigated using specific PCR followed by terminal restriction fragment length polymorphism (T-RFLP) analysis of the gene. The T-RFLP results indicated that during the period of nitrification stability, the AOB community structure in the full-scale wastewater treatment system was relatively stable, and the average change rate every 15 d of the system was 6.6%±5.8%. The phylogenetic analysis of the cloned gene showed clearly that the dominant AOB in the system was spp. The results of this study indicated that throughout the study period, the AOB community structure was relatively stable in the full-scale wastewater treatment system with functional stability of nitrification.

关键词： ammonia-oxidizing bacteria (AOB)     community dynamics     terminal restriction fragment length polymorphism (T-RFLP)     nitrification performance    

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Abundance and distribution of ammonia-oxidizing archaea in Tibetan and Yunnan plateau agricultural soils of China

Kun DING,Xianghua WEN,Liang CHEN,Daishi HUANG,Fan FEI,Yuyang LI

《环境科学与工程前沿（英文）》 2014年 第8卷 第5期   页码 693-702 doi: 10.1007/s11783-014-0635-3

摘要： As low oxygen and high ultraviolet (UV) exposure might significantly affect the microbial existence in plateau, it could lead to a specialized microbial community. To determine the abundance and distribution of ammonia-oxidizing archaea (AOA) in agricultural soil of plateau, seven soil samples were collected respectively from farmlands in Tibet and Yunnan cultivating the wheat, highland-barley, and colza, which are located at altitudes of 3200–3800 m above sea level. Quantitative PCR (q-PCR) and clone library targeting on gene were used to quantify the abundances of AOA and ammonia-oxidizing bacteria (AOB), and characterize the community structures of AOA in the samples. The number of AOA cells (9.34 × 10 –2.32 × 10 g soil) was 3.86–21.84 times greater than that of AOB cells (6.91 × 10 –1.24 × 10 g soil) in most of the samples, except a soil sample cultivating highland-barley with an AOA/AOB ratio of 0.90. Based Kendall’s correlation coefficient, no remarkable correlation between AOA abundance and the environmental factor was observed. Additionally, the diversities of AOA community were affected by total nitrogen and organic matter concentration in soils, suggesting that AOA was probably sensitive to several environmental factors, and could adjust its community structure to adapt to the environmental variation while maintaining its abundance.

关键词： ammonia-oxidizing archaea     ammonia-oxidizing bacteria     quantitative PCR     clone library     plateau    

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Interaction and independence on methane oxidation of landfill cover soil among three impact factors: water, oxygen and ammonium

Pinjing HE, Na YANG, Wenjuan FANG, Fan Lü, Liming SHAO

《环境科学与工程前沿（英文）》 2011年 第5卷 第2期   页码 175-185 doi: 10.1007/s11783-011-0320-8

摘要： To understand the influence patterns and interactions of three important environmental factors, i.e. soil water content, oxygen concentration, and ammonium addition, on methane oxidation, the soils from landfill cover layers were incubated under full factorial parameter settings. In addition to the methane oxidation rate, the quantities and community structures of methanotrophs were analyzed to determine the methane oxidation capacity of the soils. Canonical correspondence analysis was utilized to distinguish the important impact factors. Water content was found to be the most important factor influencing the methane oxidation rate and Type II methanotrophs, and the optimum value was 15% (w/w), which induced methane oxidation rates 10- and 6- times greater than those observed at 5% (w/w) and 20% (w/w), respectively. Ambient oxygen conditions were more suitable for methane oxidation than 3% oxygen. The addition of of ammonium induced different effects on methane oxidation capacity when conducted at low or high water content. With regard to the methanotrophs, Type II was sensitive to the changes of water content, while Type I was influenced by oxygen content. Furthermore, the methanotrophic acidophile, , was detected in soils with a pH of 4.9, which extended their known living environments.

关键词： quantitative polymerase chain reaction (PCR)     denaturing gradient gel electrophoresis (DGGE)     principal component analysis (PCA)     canonical correspondence analysis (CCA)    

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Assessment of antibiotic resistance genes in dialysis water treatment processes

Xuan Zhu, Chengsong Ye, Yuxin Wang, Lihua Chen, Lin Feng

《环境科学与工程前沿（英文）》 2019年 第13卷 第3期 doi: 10.1007/s11783-019-1136-1

摘要：

• Quantitative global ARGs profile in dialysis water was investigated.

• Totally 35 ARGs were found in the dialysis treatment train.

• 29 ARGs (highest) were found in carbon filtration effluent.

• erm and mtrD-02 occurred in the final effluent.

• The effluent was associated with health risks even after RO treatment.

关键词： Dialysis water     Treatment process     Antibiotic resistance gene     High-throughput quantitative PCR     Horizontal gene transfer    

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标题 作者 时间 类型 操作

662 A/G gene variation in human tumor necrosis factor receptor superfamily, member 9 (TNFRSF9)

QU Yanchun, YANG Ze, SUN Liang, JI Linong

期刊论文

Spatial and seasonal variations in bacterial communities of the Yellow Sea by T-RFLP analysis

Hongyuan WANG, Xiaolu JIANG, Ya HE, Huashi GUAN

期刊论文

Using pyrosequencing and quantitative PCR to analyze microbial communities

Husen ZHANG

期刊论文

低成本PCR微反应器阵列的温度测量与控制技术

李现明,张玉林,李建田

期刊论文

Erratum to: Using pyrosequencing and quantitative PCR to analyze microbial communities

Husen ZHANG

期刊论文

Safety evaluation of microbial pesticide (HaNPV) based on PCR method

Miao Zhao, Shufei Li, Qinghong Zhou, Dianming Zhou, Ning He, Zhiyong Qian

期刊论文

Current molecular biologic techniques for characterizing environmental microbial community

Dawen GAO, Yu TAO

期刊论文

A pooling strategy of a PCR-based assay to detect Angiostrongylus cantonensis in snail intermediate host

Fu-Rong WEI MS, Shan LV PhD, He-Xiang LIU, Ling HU, Yi ZHANG MS,

期刊论文

生物检测及分子诊断高端装备的研制及应用

彭年才,李 磊,李 政,苗保刚,李 明,赵 垚,蒋庄德

期刊论文

基于抗原和聚合酶链式反应的组合策略对模拟社区内遏制COVID-19传播效果的定量分析

黄蔷如, 孙艳侠, 贾萌萌, 张婷, 陈方圆, 姜明月, 王晴, 冯录召, 杨维中

期刊论文

半滑舌鳎促黄体激素基因克隆和表达分析及其血清浓度测定

柳学周,史宝,王珊珊,徐永江,李晓晓

期刊论文

Community dynamics of ammonia oxidizing bacteria in a full-scale wastewater treatment system with nitrification stability

Xiaohui WANG, Xianghua WEN, Hengjing YAN, Kun DING, Man HU

期刊论文

Abundance and distribution of ammonia-oxidizing archaea in Tibetan and Yunnan plateau agricultural soils of China

Kun DING,Xianghua WEN,Liang CHEN,Daishi HUANG,Fan FEI,Yuyang LI

期刊论文

Interaction and independence on methane oxidation of landfill cover soil among three impact factors: water, oxygen and ammonium

Pinjing HE, Na YANG, Wenjuan FANG, Fan Lü, Liming SHAO

期刊论文

Assessment of antibiotic resistance genes in dialysis water treatment processes

Xuan Zhu, Chengsong Ye, Yuxin Wang, Lihua Chen, Lin Feng

期刊论文