期刊文献+
共找到26篇文章
< 1 2 >
每页显示 20 50 100
Real-time fluorescent quantitative PCR非特异性扩增的研究进展 被引量:1
1
作者 刘姗姗 岳素文 +1 位作者 江洪 王成彬 《临床检验杂志(电子版)》 2013年第2期340-342,共3页
Real-time fluorescent quantitative PCR(RT-qPCR)因其高效快速、操作简便、高度敏感等优点获得广泛应用,但因其强大的放大功能而容易出现非特异性扩增的问题,尤其是荧光染料法中更加明显。文章对RT-qPCR的原理、非特异性扩增的发生因... Real-time fluorescent quantitative PCR(RT-qPCR)因其高效快速、操作简便、高度敏感等优点获得广泛应用,但因其强大的放大功能而容易出现非特异性扩增的问题,尤其是荧光染料法中更加明显。文章对RT-qPCR的原理、非特异性扩增的发生因素、解决方案及该技术的应用前景进行了综述。 展开更多
关键词 real-time fluorescent quantitative pcr 非特异性 应用
下载PDF
Application of Real-time Fluorescent Quantitative PCR in Plant
2
作者 崔颖 贾晋 +2 位作者 莎娜 李俊芳 王国泽 《Agricultural Science & Technology》 CAS 2016年第2期273-278,共6页
Real-time fluorescent quantitative PCR (RQ-PCR) is a detection method by adding fluorescent dye or fluorescent probe into the PCR reaction system, using fluorescent signal accumulation to monitor amplification react... Real-time fluorescent quantitative PCR (RQ-PCR) is a detection method by adding fluorescent dye or fluorescent probe into the PCR reaction system, using fluorescent signal accumulation to monitor amplification reactions of PCR reaction process, and finally the unknown template can be quantitatively analyzed through the standard curve. So the detection level of PCR has improved from the qualitative to the quantitative. In order to provide a theoretical reference for further application, the principle, classification, advantages and disadvantages of RQ-PCR were intro- duced, and its application and progress in plants in recent years were reviewed. 展开更多
关键词 real-time fluorescent quantitative pcr (RQ-pcr PRINCIPLE Reference gene Stress resistance of plant Transgenic product
下载PDF
Detection of Lactobacillus acidophilus in Fermented Material by Real-time Fluorescent Quantitative PCR 被引量:4
3
作者 Guo Zihao Fang Hua +4 位作者 Xia Zhisheng Zhu Xiaoshi Sun Zhongchao Yu Hanli Xia Jiaji 《Animal Husbandry and Feed Science》 CAS 2016年第1期54-57,共4页
The species distinctive PCR primer of Lactobacillus acidophilus ( L. acidophilus) was designed according to 16S rRNA gene sequences of conunon Lac- tobacillus species in fermented material. Bacterial genome DNA of s... The species distinctive PCR primer of Lactobacillus acidophilus ( L. acidophilus) was designed according to 16S rRNA gene sequences of conunon Lac- tobacillus species in fermented material. Bacterial genome DNA of separated L. acidophilus in fermented sample was taken as template, and L. acidophilus in fer- mented material was conducted the quantitative determination by real-time quantitative PCR (RT-PCR). Analysis on RT-PCR results shown that contents of L. aci- dophilus in the test sample reached 1.5 billion CFU / g. Test results shown that contents of L. acidophilus in fermented material could be detected accurately by the established RT-PCR method in the test. indicating that the established RT-PCR method could be aookued to the detection of L. acidophilus in fermented material. 展开更多
关键词 real-time fluorescent quantitative pcr Lactobacillus acidophilus quantitative analysis Fermented material
下载PDF
Application of Real-time Fluorescent Quantitative PCR in Studies on Plants 被引量:3
4
作者 Yueping MA Silan DAI Yanrong MA 《Agricultural Biotechnology》 CAS 2012年第1期1-7,共7页
Real-Lime fluorescent quantitative PCR is a method for quantitative analysis of gene expression developed in recent years, which has been widely used in various fields such as basic scientific research, clinical diagn... Real-Lime fluorescent quantitative PCR is a method for quantitative analysis of gene expression developed in recent years, which has been widely used in various fields such as basic scientific research, clinical diagnosis, disease study, drug research and development since its appearance. It starts relatively late in study on plants, but has already been used for analysis of gene expression in plants and gene identification of exogenous genes. The principles or advantages and dis- advantages of real-time fluorescent quantitative PCR, or its potential problems and condition optimizations in tests were introduced in this study, and then the appli- cation and prospect of real-time fluorescent quantitative PCR in study on plants were also been discussed. 展开更多
关键词 real-time fluorescent quantitative pcr (FQ-pcr PLANT C ene expression
下载PDF
Establishment of a Real-time Fluorescent Quantitative PCR Assay for Detection of Genetically Modified Maize Line MON88017
5
作者 Jun SONG Dong WANG 《Agricultural Biotechnology》 CAS 2017年第1期15-19,22,共6页
In order to improve the standardized technical systems of quantitative analyses for genetically modified organisms (GMOs) and products, ensure bio-safety and reduce ecological risk in China, a real-time fluorescent ... In order to improve the standardized technical systems of quantitative analyses for genetically modified organisms (GMOs) and products, ensure bio-safety and reduce ecological risk in China, a real-time fluorescent quantitative PCR assay was established for detection of genetically modified maize line MON88017. The established method was evaluated based on the specificity, sensitivity, accuracy and measurement uncertainty. The results showed that the established method had strong specificity in detection of genetically modified maize line MON88017. 1.50% MON88017 sample was detected with 29 replica- tions. The average measured value ( 1. 541% ) was close to the actual value ( 1.50% ) and the relative deviation was 2.70%. The variation coefficient of the measured value was 0.110 g ; the recovery was 100.00% and the measurement uncertainty was 0. 096. The limit of detection for genetically modified maize line MON88017 with the established method was 5 copies at the 97.5% confidence level. Thus, the real-time fluorescent quantitative PCR assay established in this study exhibited high specificity, accuracy and sensitivity, which could provide technical support for the safety supervision of genetically modified organ- isms and products in China. 展开更多
关键词 Genetically modified maize real-time fluorescent quantitative pcr SPECIFICITY Sensitivity ACCURACY Measurement uncertainty
下载PDF
Detection of Ratoon Stunting Disease in Virus-free Seedcane via Real-time Fluorescence Quantitative PCR 被引量:1
6
作者 Ming DAN Song LI +3 位作者 Kunxing YU Limin LIU Hongjian LIU Manman LU 《Agricultural Biotechnology》 CAS 2012年第5期24-26,共3页
This study was to develop the real-time fluorescence quantitative PCR technique for detecting the ratoon stunting disease (RSD) in virus-free seedcane seedlings. Healthy tissue culture seedlings were obtained from s... This study was to develop the real-time fluorescence quantitative PCR technique for detecting the ratoon stunting disease (RSD) in virus-free seedcane seedlings. Healthy tissue culture seedlings were obtained from six plants of sugarcane ROC22, which had been confirmed RSD-positive by detecting the sugarcane juice, by employing the sugarcane seedlings production protocol. Real-time fluorescence quantitative PCR was used to detect RSD pathogens in tissue culture sam- pies. The results showed that target fragment of RSD pathogens was not found in all 10 samples in real-time fluorescence quantitative PCR, with the Ct values of 37 - 39. The healthy tissue culture sugarcane seedlings do not carry RSD pathogens, indicating that adopting healthy seedcane seedlings production technique could thoroughly get rid of RSD pathogens. 展开更多
关键词 SUGARCANE Virus-free seedcane Ratoon stunting disease real-time fluorescence quantitative pcr
下载PDF
Development and Preliminary Application of SYBR Green I Real-Time Fluorescence Quantitative PCR Method for Detecting Porcine Parvovirus Virus
7
作者 SHEN Zhi-qiang WANG Jin-liang +3 位作者 GUO Xian-po WANG Xiao-hu WANG Ming ZHAO De-ming 《Animal Husbandry and Feed Science》 CAS 2009年第11期42-46,共5页
According to VP2 gene sequence of the porcine parvovirus virus strain NADL-2 (NC001718) available in GenBank (NC_001718), a pair of specific primer was designed, and the target fragment of 431 bp was obtained by P... According to VP2 gene sequence of the porcine parvovirus virus strain NADL-2 (NC001718) available in GenBank (NC_001718), a pair of specific primer was designed, and the target fragment of 431 bp was obtained by PCR amplification. The products were ligated with pMD18- T vector and then transformed into bacteria DH5α for recombinant plasmid extraction. After PCR identification and sequencing, recombinant plasmid was used as a standard template to establish the standard curve of SYBR Green I fluorescence quantitative PCR. Sensitivity test, specificity test and repeatability test were also determined. The results indicated that there was a good linear relationship between threshold cycle of the standard curve and template concentration, R2 =0.997 6. Tm ranged from 82.3 to 82.9 ℃, while the sensitivity was 72.1 copies/μl with good specificity and repeatability. The developed SYBR Green I real-time quantitative PCR method to detect PPV VP2 gene laid the basis for further studies on patho- oenesis, early clinical diaonosis of this virus and quantitative analysis of PPV infection. 展开更多
关键词 Porcine parvovirus virus real-time fluorescence quantitative pcr DETECTION
下载PDF
Primary application of a real-time quantitative polymerase chain reaction for the detection of human breast cancer related novel gene-Metadherin expression 被引量:1
8
作者 Bing Li Zhaozhe Liu Xiaodong Xie Yakun Wang 《The Chinese-German Journal of Clinical Oncology》 CAS 2010年第6期316-320,共5页
Objective:The aim of this study was to detect the expression level of Metadherin (MTDH) in peripheral blood of the breast cancer patients by real-time fluorescence quantitative polymerase chain reaction (PCR),and to e... Objective:The aim of this study was to detect the expression level of Metadherin (MTDH) in peripheral blood of the breast cancer patients by real-time fluorescence quantitative polymerase chain reaction (PCR),and to explore the relationship between expression of Metadherin gene in the patients peripheral blood and the clinic-pathological features in breast cancer. Methods:Real-time fluorescence quantitative polymerase chain reaction was employed to determine the expression level of Metadherin gene in 80 peripheral blood samples of breast cancer patients and healthy donors. Results:The expression of Metadherin gene in breast cancer patients peripheral blood were positive,in which 34 breast cancer patients were highly expressed,accounting for 55.7%,while the expression of Metadherin gene in normal females peripheral blood were negative,there was statistical significance (Ratio = 2.02±0.81,P < 0.05); Ratio of the Metadherin expression in breast cancer patients peripheral blood and the glyceraldehyde-3-phosphate dehydrogenase expression was 1.15 ± 0.36. REST software analysis showed that the expression of Metadherin gene was significantly up-regulated in breast cancer. Conclusion:The SYBR Green I quantitative real-time polymerase chain reaction method can successfully detect the expression level of Metadherin gene. Expression level of Metadherin gene in breast cancer patients peripheral blood is closely related to survival,and it maybe involved in the development of breast cancer and used as an indicator of prognosis. 展开更多
关键词 breast cancer Metadherin (MTDH) real-time fluorescence quantitative polymerase chain reaction pcr
下载PDF
Development of real-time PCR method for rapid detection and quantification of Heterosigma akashiwo 被引量:1
9
作者 何闪英 于志刚 米铁柱 《Journal of Harbin Institute of Technology(New Series)》 EI CAS 2008年第1期118-123,共6页
To rapidly detect the harmful algae H.akashiwo qualitatively and quantitatively, sequences of the 18S rDNA deduced from H.akashiwo were used for designing species-specific primers, and a RFQ-PCR (Real-time Fluorescent... To rapidly detect the harmful algae H.akashiwo qualitatively and quantitatively, sequences of the 18S rDNA deduced from H.akashiwo were used for designing species-specific primers, and a RFQ-PCR (Real-time Fluorescent Quantitative Polymerase Chain Reaction) method was developed for quantitative detection of H.akashiwo. Primer H.akashiwo and TaqMan probe were designed, and the specificity of primer was checked with PCR. A calibration curve was constructed with cycle threshold value against visual counted cell number. And the value of the curve was tested with other H.akashiwo samples, which were assayed with both the RFQ-PCR method and visual count under microscope. 展开更多
关键词 Heterosigma akashiwo fluorescent quantitative pcr molecular probe real-time detection
下载PDF
ABI 7300实时荧光定量PCR仪的性能评估 被引量:6
10
作者 巢薇 《国际检验医学杂志》 CAS 2015年第4期494-495,共2页
目的为参加医学实验室认可和评价仪器性能,对ABI 7300实时荧光定量PCR仪的性能进行评估。方法参照美国临床实验室标准化委员会(CLSI)文件对ABI 7300荧光定量PCR仪检测HBV-DNA定量结果的精密度、正确度、灵敏度及线性进行评价。结果精... 目的为参加医学实验室认可和评价仪器性能,对ABI 7300实时荧光定量PCR仪的性能进行评估。方法参照美国临床实验室标准化委员会(CLSI)文件对ABI 7300荧光定量PCR仪检测HBV-DNA定量结果的精密度、正确度、灵敏度及线性进行评价。结果精密度:批内变异系数(CV批内)为2.38%-4.61%,批间变异系数(CV批间)为3.87%-5.33%,达到《医学实验室质量和能力认可准则》中基因扩增检验项目分析性能标准(CV批内〈4.8%,CV批间〈6.4%)。正确度:参加2011年卫生部室间质评结果总体平均偏倚为2.14%,符合卫生部质评要求(偏倚不超过8%);功能灵敏度:100IU/mL检测限浓度的CV值最接近于20%,与试剂盒检出下限相符;线性:相关系数(r2)为1.0,线性关系符合要求。结论 ABI 7300实时荧光定量PCR仪各项性能指标精确,可以为临床提供快速、准确的报告。 展开更多
关键词 实时荧光定量pcr 性能评估 HBV-DNA
下载PDF
定量PCR分析仪的临床应用前景——AG-9900 Robo Master全自动荧光定量PCR分析仪
11
作者 毛远丽 《引进国外医药技术与设备》 1999年第11期60-61,共2页
荧光探针杂交定量PCR技术和全自动定量PCR分析仪的出现,使PCR扩增和产物分析整个过程完全在同一个封闭系统下完成,并在分析软件支持下实现对PCR扩增产物进行实时动态监测和自动得出定量结果。彻底解决了PCR技术中扩增产物污染和不能定... 荧光探针杂交定量PCR技术和全自动定量PCR分析仪的出现,使PCR扩增和产物分析整个过程完全在同一个封闭系统下完成,并在分析软件支持下实现对PCR扩增产物进行实时动态监测和自动得出定量结果。彻底解决了PCR技术中扩增产物污染和不能定量的问题,为PCR技术作为常规检测技术应用于临床开辟了广阔的前景。 展开更多
关键词 聚合酶链反应 pcr分析仪 荧光定量分析
下载PDF
Analysis of Seed-specificity of Silencing fad_2 Gene Expression in Transgenic Rapeseed Line W-4(Brassica napus L.) 被引量:3
12
作者 陈松 彭琦 +5 位作者 周晓婴 高建芹 张维 张洁夫 浦惠明 戚存扣 《Agricultural Science & Technology》 CAS 2014年第8期1308-1311,1316,共5页
This study was to investigate the efficiency and specificity of RNAi silencing on the expression of endogenous fad2 gene in transgenic line W-4. [Method] The relative expression of fad2 gene in seeds at different deve... This study was to investigate the efficiency and specificity of RNAi silencing on the expression of endogenous fad2 gene in transgenic line W-4. [Method] The relative expression of fad2 gene in seeds at different developmental stages of 7th, 14th, 21st and 28th day after flowering (DAF) as wel as the root, stem, leaf at winter seedling stages of both the transgenic line W-4 and non-transgenic control Westar by real-time fluorescence quantitative PCR. [Results] The results showed the relative expression of fad2 gene was gradual y increasing with the days after flowering in the seeds of the control Westar, while it was found decreasing significantly since the 21st DAF in the seeds of the line W-4. The decline was up to 60% in comparison with the control Westar. However, no significant difference in the relative expression of fad2 gene in other organs like root, stem and leaf was observed between transgenic line W-4 and non-transgenic control Westar. Fatty acid composition analysis showed the oleic acid desaturation parameter(ODP) in seeds of the line W-4 was 0.07 in average, decreased by nearly 75% than control Westar which was 0.24 in average, while no significant difference in the seedling root, stem and leaf was measured between transgenic rapeseed and control. [Conclusion] The results above validated that RNA interference in transgenic rapeseed W-4 is at a seed-specific manner, not interfering with fad2 gene expression in organs such as the root, stem and leaf. The study also found that the period of fad2 gene expres-sion decline was wel coincided with the expression of napin gene, both appeared at the 21st DAF, indicating that the expression of dsRNA of fad2 gene is precisely control ed by the napin promoter. 展开更多
关键词 Transgenic rapeseed real-time fluorescence quantitative pcr fad2gene Specific expression
下载PDF
Changes of the Transcriptional Levels of Molecules Associated with Endogenous Antigen Processing and Presentation in Porcine Skin-derived Dendritic Cells Infected with PCV2 in vivo 被引量:1
13
作者 李建东 李焕荣 +2 位作者 聂晓华 遇奇 崔德凤 《Agricultural Science & Technology》 CAS 2012年第5期1089-1092,共4页
[Objective] This study aimed to investigate the changes of the transcriptional levels of molecules associated with endogenous antigen processing and presenta- tion in porcine skin-derived dendritic cells infected with... [Objective] This study aimed to investigate the changes of the transcriptional levels of molecules associated with endogenous antigen processing and presenta- tion in porcine skin-derived dendritic cells infected with PCV2 in vivo. [Method] Healthy 40-day-old Landrace piglets were infected with porcine circovirus type 2 (PCV2) and euthanized on the 34, 7rd, 14th, 21st and 35th d post inoculation (DPI). The porcine skin-derived dendritic cells (DCs) were collected to analyze the transcrip- tional levels of molecules (LMP7, UBP, MHC-I, calreticulin) associated with endogenous antigen processing and presentation by using real-time fluorescent quantitative PCR (real-time FQ-PCR). [Result] The results showed that the level of LMP7 mR- NAs was reduced significantly on the 3DPI (P〈0.05); the level of UBP mRNAs was consistently up-regulated, which increased significantly on the 21DPI and 35DPI (P〈 0.05); the level of MHC-I mRNAs was significantly down-regulated on the 7DPI (P〈 0.05); the level of calreticulin mRNAs was up-regulated slightly without significant dif- ference. [Conclusion] PCV2 can inhibit the endogenous antigen processing and presentation ability of porcine skin-derived DCs at early stages of infection. 展开更多
关键词 Porcine circovirus type 2 Skin-derived dendritic cells Endogenous antigen processing and presentation real-time fluorescent quantitative pcr
下载PDF
MecA、Nuc基因在耐甲氧西林金黄色葡萄球菌检测中的应用 被引量:3
14
作者 郭建巍 齐文峰 +6 位作者 郝秀红 李文军 陈昌国 张云 马志家 陈秋圆 赵强元 《转化医学杂志》 2017年第3期157-159,共3页
目的通过对180株金黄色葡萄球菌临床分离株MecA、Nuc基因的检测,以期选择出一种适合临床的并能简便、快速、准确检测耐甲氧西林金黄色葡萄球菌(methicillin resistant Staphylococcus aureus,MRSA)的方法。方法细菌鉴定及药敏采用全自... 目的通过对180株金黄色葡萄球菌临床分离株MecA、Nuc基因的检测,以期选择出一种适合临床的并能简便、快速、准确检测耐甲氧西林金黄色葡萄球菌(methicillin resistant Staphylococcus aureus,MRSA)的方法。方法细菌鉴定及药敏采用全自动细菌鉴定和药敏分析仪。MecA、Nuc基因检测采用荧光定量聚合酶链反应(fluorescence quantitative polymerase chain reaction,FQ-PCR)法。结果 FQ-PCR对180株金黄色葡萄球菌MecA、Nuc基因检出率为75.0%,全自动细菌鉴定和药敏分析仪对180株金黄色葡萄球菌的MRSA检出率为72.8%,2种检测方法比较差异无统计学意义(P>0.05)。全自动细菌鉴定和药敏分析仪对MRSA的检测正确率为94.8%。结论 FQ-PCR检测MRSA正确率高,只需要1~2 h即可完成。该方法快速、简便、准确,值得推广应用。 展开更多
关键词 耐甲氧西林金黄色葡萄球菌 MECA基因 Nuc基因 荧光定量聚合酶链反应 全自动细菌鉴定和药敏分析仪
下载PDF
Analysis of Gene Expression of Seven Isoforms of ADP-glucose Pyrophosphorylase in Rice Endosperm under Different Temperature Conditions
15
作者 袁定阳 孙志忠 +1 位作者 谭炎宁 段美娟 《Agricultural Science & Technology》 CAS 2012年第6期1226-1229,1233,共5页
[Objective] This study aimed to analyze the effects of temperature on the expression of AGPase isoform genes in rice endosperm during milk stage. [Method] Different temperature treatments (33 and 25 ℃ of daily mean ... [Objective] This study aimed to analyze the effects of temperature on the expression of AGPase isoform genes in rice endosperm during milk stage. [Method] Different temperature treatments (33 and 25 ℃ of daily mean temperature for high and normal temperature treatments, respectively) and the real-time fluorescence quantitative PCR ( FQPCR) were used to analyze the expression patterns of seven isoforms (AGPS1, AGPS2a, AGPS2b, AGPL1, AGPL2, AGPL3 and AGPL4) of ADPglucose pyrophosphorylase (AGPase) which was the key enzyme in starch synthesis and metabolism in rice endosperm of two rice varieties Teqing and Thai Fragrant Rice. [Result] The AGPase isoforms AGPS2b, AGPL2 and AGPL3 had much higher expression than the other four isoforms, thus they were thought to be the main expression patterns of AGPase in rice endosperm. The relative expressions of AGPL2 was the highest among all the isoforms. The relative expressions of AGPS2b, AGPL2 and AGPL3 were higher in the normal temperature treatment than in the high temperature treatment in both rice varieties. The relative expression of the three enzyme genes in milk stages in Teqing was higher than those in Thai Fragrant Rice under different temperature treatments. [Conclusion] This study provides a theoretical basis for further use of molecular biology techniques to cultivate stable high-quality rice varieties. 展开更多
关键词 RICE ADP-glucose pyrophosphorylase (AGPase) isoforms Gene expression characteristics real-time fluorescence quantitative pcr (FQ-pcr
下载PDF
Analysis on Tissue-specific Expression of Dp XTH1 and Dp XTH2 Genes in Dahlia 被引量:1
16
作者 张萍萍 王蕾 +4 位作者 陈驰 王利芬 郑必平 钱力鑫 谈建中 《Agricultural Science & Technology》 CAS 2015年第8期1596-1599,共4页
[Objective] This study aimed to investigate the functions and related mechanisms of xyloglucan Endotransglycosylase/hydrolases (XTHs) during the growth and development of dahlia. [Method] Using /3-actin as the refer... [Objective] This study aimed to investigate the functions and related mechanisms of xyloglucan Endotransglycosylase/hydrolases (XTHs) during the growth and development of dahlia. [Method] Using /3-actin as the reference gene, the rela- tive transcription levels of DpXTH1 and DpXTH2 genes in roots, stems, leaves and petals of dahlia were analyzed by real-time RT-PCR. [Result] The DpXTH1 and DpXTH2 were not expressed in the roots, but expressed abundantly in the petals of dahlia. There were little expressions in the stems and leaves of dahlia. [Conclusion] The DpXTH1 and DpXTH2 were petal-specific genes and closely related to the growth and development of petals in dahlia. 展开更多
关键词 DAHLIA Xyloglucan endotransglycosylase/hydrolases XTH gene Expres-sion specificity real-time fluorescence quantitative pcr
下载PDF
Efflux pump gene hefA of Helicobacter pylori plays an important role in multidrug resistance 被引量:17
17
作者 Zhi-Qiang Liu Peng-Yuan Zheng Ping-Chang Yang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2008年第33期5217-5222,共6页
AIM: To determine whether efflux systems contribute to multidrug resistance of H pylori. METHODS: A chloramphenicol-induced multidrug resistance model of six susceptible H pylori strains (5 isolates and H pylori NCTC1... AIM: To determine whether efflux systems contribute to multidrug resistance of H pylori. METHODS: A chloramphenicol-induced multidrug resistance model of six susceptible H pylori strains (5 isolates and H pylori NCTC11637) was developed. Multidrug-resistant (MDR) strains were selected and the minimal inhibitory concentration (MIC) of eryth-romycin, metronidazole, penicillin G, tetracycline, and ciprofloxacin in multidrug resistant strains and their parent strains was determined by agar dilution tests. The level of mRNA expression of hefA was assessed by fluorescence real-time quantitative PCR. A H pylori LZ1026 knockout mutant (ΔH pylori LZ1026) for (puta-tive) efflux protein was constructed by inserting the kanamycin resistance cassette from pEGFP-N2 into hefA, and its susceptibility profiles to 10 antibiotics were evaluated. RESULTS: The MIC of six multidrug-resistant strains (including 5 clinical isolates and H pylori NCTC11637) increased signifi cantly (≥ 4-fold) compared with their parent strains. The expression level of hefA gene was significantly higher in the MDR strains than in their parent strains (P = 0.033). A H pylori LZ1026 mutant was successfully constructed and the ΔH pylori LZ1026 was more susceptible to four of the 10 antibiotics. All the 20 strains displayed transcripts for hefA that con-fi rmed the in vitro expression of these genes.CONCLUSION: The efflux pump gene hefA plays an important role in multidrug resistance of H pylori. 展开更多
关键词 Efflux pump Helicobacter pylori Multidrug resistance fluorescence real-time quantitative pcr Knockout mutant
下载PDF
Temperature Stress at Grain Filling Stage Mediates Expression of Three Isoform Genes Encoding Starch Branching Enzymes in Rice Endosperm 被引量:3
18
作者 WEI Ke-su CHENG Fang-min ZHANG Qi-fang Liu Kui-gang 《Rice science》 SCIE 2009年第3期187-193,共7页
An early-maturity indica rice variety Zhefu 49, whose grain quality and starch structure are sensitive to environmental temperature, was subjected to different temperatures (32℃ for high temperature and 22℃ for opt... An early-maturity indica rice variety Zhefu 49, whose grain quality and starch structure are sensitive to environmental temperature, was subjected to different temperatures (32℃ for high temperature and 22℃ for optimum temperature) at the grain filling stage in plant growth chambers, and the different expressions of three isoform genes (SBEI, SBEIII and SBE/V) encoding starch branching enzyme (SBE) in the endosperms were studied by the real-time fluorescence quantitative PCR (FQ-PCR) method. Effects of high temperature on the SBE expression in developing rice endosperrns were isoform-dependent. High temperature significantly down-regulated the expressions of SBEI and SBEIII, while up-regulated the expression of SBEIV. Compared with SBEIV and SBEIII, the expression of SBEI gene in Zhefu 49 rice endosperms was more sensitive to temperature variation at the grain filling stage. This study indicates that changes in weather/climate conditions especially temperature stress influence rice grain formation and its quality as evidenced by isoform expression. 展开更多
关键词 RICE high temperature starch branching enzyme ISOFORM gene expression real-time fluorescence quantitative pcr rice quality
下载PDF
Expressions of Toll Like Receptor(TLR)Genes in Paralichthys olivaceus After Induced by Different Extracts of Edwardsiella tarda 被引量:2
19
作者 SHAN Yanan ZHENG Jinhui +1 位作者 GAO Hong SUN Jinsheng 《Journal of Ocean University of China》 SCIE CAS CSCD 2022年第4期1027-1036,共10页
Toll like receptors(TLRs)are the main innate immune‘pattern recognition receptors’of animals,which play a central role in host cell recognition and responses to invasive pathogens,particularly common structures of m... Toll like receptors(TLRs)are the main innate immune‘pattern recognition receptors’of animals,which play a central role in host cell recognition and responses to invasive pathogens,particularly common structures of microbial pathogens.In this study,the gene expression profiles of TLRs in the spleen,head kidney,gill,small intestine,liver,muscle,and heart of healthy Paralichthys olivaceus were detected by real-time quantitative PCR(qPCR).The TLR family members were widely expressed in different tissues with different basic expression profiles.The highest expressions of TLR1,5m,7,8,9,14,and 21 were found in the spleen;the highest expressions of TLR3 and TLR21 were found in the gill;the highest expressions of TLR2 and 5s were found in the small intestine.The second highest expressions of TLR3,7,and 8 were found in small intestine.The gene expression profiles of TLRs stimulated with Edwardsiella tarda DNA,RNA,and lipopolysaccharide(LPS)were also detected in spleen,head kidney and gill.TLR9 and TLR21 were sensitive to E.tarda DNA;TLR 8 and TLR21 were sensitive to E.tarda RNA;and TLR1 and TLR14 were sensitive to E.tarda LPS.The expressions of the other TLR genes showed no significant changes.The results imply that the expressions of these TLR genes in P.olivaceus are differently regulated in the whole body and play important roles in the immune response against E.tarda infection. 展开更多
关键词 Edwardsiella tarda Paralichthys olivaceus TLRS EXPRESSION real-time fluorescence quantitative pcr
下载PDF
全自动化学发光分析仪及核酸扩增仪检测乙肝标志物的应用 被引量:6
20
作者 李林臣 张懿晖 +2 位作者 牛艳芬 李芙琴 谢桂芳 《中国医学装备》 2016年第4期41-44,共4页
目的:探讨全自动化学发光分析仪及核酸扩增仪检测乙肝标志物的临床应用。方法:选择2235例住院及门诊收治的肝功能异常病例,分别用化学发光方法检测乙肝病毒表面抗原(HBs Ag),用实时荧光聚合酶链式反应(FQ-PCR)方法检测乙肝病毒基因(HBV-... 目的:探讨全自动化学发光分析仪及核酸扩增仪检测乙肝标志物的临床应用。方法:选择2235例住院及门诊收治的肝功能异常病例,分别用化学发光方法检测乙肝病毒表面抗原(HBs Ag),用实时荧光聚合酶链式反应(FQ-PCR)方法检测乙肝病毒基因(HBV-DNA)含量,并对结果进行分析。每位患者检测HBs Ag与HBV-DNA均采用同一份标本。结果:全部受检标本中,化学发光方法检测HBs Ag阳性率为61.34%,显著高于FQ-PCR方法检测阳性率(48.59%),差异有统计学意义(x^2=73.41,P<0.01)。将化学发光检测HBs Ag的结果分成强阳性、弱阳性和阴性3组,不同组间FQ-PCR检测HBV-DNA的阳性率显著不同,差异有统计学意义(x^2=1863.60,P<0.01),强阳性组HBV-DNA阳性率为97.31%,显著高于弱阳性组,差异有统计学意义(x^2=966.90,P<0.01)。结论:全自动化学发光分析仪所采用的化学发光方法对筛查乙肝病毒感染效果较好,而扩增仪所采用的FQ-PCR方法对检测乙肝病毒复制及疗效的监测效果更好,二者联合检测可提高乙肝患者的检出率并可对乙肝患者的病情及疗效作出准确判断。 展开更多
关键词 全自动化学发光仪 聚合酶链式反应 扩增仪 乙肝表面抗原 乙型肝炎病毒脱氧核糖核酸
下载PDF
上一页 1 2 下一页 到第
使用帮助 返回顶部