资源类型

期刊论文 19

年份

2023 1

2019 3

2018 2

2016 1

2015 1

2014 2

2013 1

2012 1

2011 4

2010 1

2008 1

2007 1

展开 ︾

关键词

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

聚合酶链反应(PCR) 1

聚合酶链式反应 1

展开 ︾

检索范围:

排序: 展示方式:

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    

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    

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

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    

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    

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    

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    

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    

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    

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)    

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    

Seasonal microbial community shift in a saline sewage treatment plant

Qingmei YAN, Xuxiang ZHANG, Tong ZHANG, Herbert H P FANG

《环境科学与工程前沿(英文)》 2011年 第5卷 第1期   页码 40-47 doi: 10.1007/s11783-011-0304-8

摘要: Activated sludge was monthly sampled from a saline sewage treatment plant of Hong Kong (China) during June 2007 to May 2008 to analyze the microbial community shift along with environmental variations using denaturing gradient gel electrophoresis of polymerase chain reaction amplified 16S rDNA fragments. Environmental changes resulted into a seasonal microbial community shift characterized by alterations in species number and abundance in the sewage treatment plant. Correspondence analysis and cluster analysis on community structure profile showed that the 12 monthly samples fell into four groups, which is in accordance with season changing in Hong Kong. Canonical correspondence analysis revealed that and posed more significant effects on community structure than total phosphorus and total nitrogen, respectively. Compared with sludge retention time, influent total phosphorus had an inverse effect on the community structure shift, and chemical oxygen demand and showed a similar effects. Results of this study may contribute to the development of new knowledge involving the microbial community shift in sewage treatment plants.

关键词: sewage treatment plants     polymerase chain reaction (PCR)     denaturing gradient gel electrophoresis (DGGE)     canonical correspondence analysis     correspondence analysis    

Nucleic acid-based diagnostics for infectious diseases in public health affairs

null

《医学前沿(英文)》 2012年 第6卷 第2期   页码 173-186 doi: 10.1007/s11684-012-0195-5

摘要:

Infectious diseases, mostly caused by bacteria and viruses but also a result of fungal and parasitic infection, have been one of the most important public health concerns throughout human history. The first step in combating these pathogens is to get a timely and accurate diagnosis at an affordable cost. Many kinds of diagnostics have been developed, such as pathogen culture, biochemical tests and serological tests, to help detect and fight against the causative agents of diseases. However, these diagnostic tests are generally unsatisfactory because they are not particularly sensitive and specific and are unable to deliver speedy results. Nucleic acid-based diagnostics, detecting pathogens through the identification of their genomic sequences, have shown promise to overcome the above limitations and become more widely adopted in clinical tests. Here we review some of the most popular nucleic acid-based diagnostics and focus on their adaptability and applicability to routine clinical usage. We also compare and contrast the characteristics of different types of nucleic acid-based diagnostics.

关键词: nucleic acid-based diagnostics     infectious disease     PCR     NASBA     LAMP     microarray     LOAC     public health affairs    

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    

Simultaneous detection and characterization of toxigenic:Clostridium difficile directly from clinical stool specimens

null

《医学前沿(英文)》 2018年 第12卷 第2期   页码 196-205 doi: 10.1007/s11684-017-0560-5

摘要:

We employed a multiplex polymerase chain reaction (PCR) coupled with capillary electrophoresis (mPCR-CE) targeting six Clostridium difficile genes, including tpitcdAtcdBcdtAcdtB, and a deletion in tcdC for simultaneous detection and characterization of toxigenic C. difficile directly from fecal specimens. The mPCR-CE had a limit of detection of 10 colony-forming units per reaction with no cross-reactions with other related bacterial genes. Clinical validation was performed on 354 consecutively collected stool specimens from patients with suspected C. difficile infection and 45 isolates. The results were compared with a reference standard combined with BD MAX Cdiff, real-time cell analysis assay (RTCA), and mPCR-CE. The toxigenic C. difficile species were detected in 36 isolates and 45 stool specimens by the mPCR-CE, which provided a positive rate of 20.3% (81/399). The mPCR-CE had a specificity of 97.2% and a sensitivity of 96.0%, which was higher than RTCA (x2 = 5.67, P = 0.017) but lower than BD MAX Cdiff (P = 0.245). Among the 45 strains, 44 (97.8%) were determined as non-ribotype 027 by the mPCR-CE, which was fully agreed with PCR ribotyping. Even though ribotypes 017 (n = 8, 17.8%), 001 (n = 6, 13.3%), and 012 (n = 7, 15.6%) were predominant in this region, ribotype 027 was an important genotype monitored routinely. The mPCR-CE provided an alternative diagnosis tool for the simultaneous detection of toxigenic C. difficile in stool and potentially differentiated between RT027 and non-RT027.

关键词: Clostridium difficile     multiplex PCR     capillary electrophoresis     detection     characterization    

标题 作者 时间 类型 操作

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

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

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

Seasonal microbial community shift in a saline sewage treatment plant

Qingmei YAN, Xuxiang ZHANG, Tong ZHANG, Herbert H P FANG

期刊论文

Nucleic acid-based diagnostics for infectious diseases in public health affairs

null

期刊论文

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

Simultaneous detection and characterization of toxigenic:Clostridium difficile directly from clinical stool specimens

null

期刊论文