目的建立荧光定量PCR检测美洲钩虫的方法。方法提取虫体基因组DNA、根据GenBank中美洲钩虫ITS-2序列设计特异引物,PCR扩增ITS-2序列、克隆、测序和比对。常规PCR检验引物特异性。将ITS-2序列扩增产物回收、纯化后经T克隆转入大肠埃希菌...目的建立荧光定量PCR检测美洲钩虫的方法。方法提取虫体基因组DNA、根据GenBank中美洲钩虫ITS-2序列设计特异引物,PCR扩增ITS-2序列、克隆、测序和比对。常规PCR检验引物特异性。将ITS-2序列扩增产物回收、纯化后经T克隆转入大肠埃希菌DH5α,提取重组质粒,鉴定后作为标准品模板建立荧光定量PCR标准曲线,并做灵敏性和重复性试验。结果构建的荧光定量PCR标准曲线循环阈值与模板浓度呈良好的线性关系,溶解曲线的吸收峰单一,实验重复性良好。结论成功构建了SYBR Green Ⅰ荧光定量PCR检测美洲钩虫的方法,可用于快速、准确、定量检测美洲钩虫。展开更多
目前针对胡适少年时期思想的研究,通常使用的是胡适的《四十自述》及《我的信仰》(What I Believe)。论文通过《胡适日记》及书信的记载,考察胡适的My Credo and Its Evolution与What I Believe的内容与成书时间,提出它们就是写作《四...目前针对胡适少年时期思想的研究,通常使用的是胡适的《四十自述》及《我的信仰》(What I Believe)。论文通过《胡适日记》及书信的记载,考察胡适的My Credo and Its Evolution与What I Believe的内容与成书时间,提出它们就是写作《四十自述》的最初蓝本。展开更多
This paper provides a critical analysis of the design and implementation of Graduate English 1 for 1st year post graduates,9 credits to gain toward the degree.Specifically,the paper examines the details of the module ...This paper provides a critical analysis of the design and implementation of Graduate English 1 for 1st year post graduates,9 credits to gain toward the degree.Specifically,the paper examines the details of the module specification template(MST) and provides an analysis on the aims,objectives,teaching methods,learning outcomes and assessment within the learning program,with reference to current theory on curriculum design and assessment.展开更多
Currently, available phenotyping and commercial methods report <em>A. baumannii</em> only as <em>Acinetobacter calcoaceticus-baumannii</em> complex (ACB) and do not identify individual members ...Currently, available phenotyping and commercial methods report <em>A. baumannii</em> only as <em>Acinetobacter calcoaceticus-baumannii</em> complex (ACB) and do not identify individual members of the complex. This is a single blind study aimed to evaluate certain commonly used species-specific genetic markers namely, Intergenic Transcribed Spacer region in 16S rRNA of <em>A. baumannii</em> (Ab-ITS) and <em>gyrB</em>, for identification of ACB members. These molecular targets were first validated on clinical isolates (n = 200) and subsequently on uncultured tracheal aspirates (n = 172). Among the clinical isolates, 183/200 (91.5%) were positive for Ab-ITS. The clinical isolates 17 (17/200) which are failed to amplify in Ab-ITS PCR were subsequently diagnosed by <em>gyrB</em> PCR as <em>A. calcoaceticus</em> (n = 2), <em>A. pitti</em> (n = 6) and <em>A. nosocomialis</em> (n = 9) but not <em>A. baumannii</em>. Among the tracheal aspirates, 62 samples were reported as sterile in Advanced Expert System of VITEK-2, among the remaining 110 samples, 68.1% (75/110) samples contained Ab-ITS target. Twenty-five of the sterile samples (25/62) were found to contain Ab-ITS target sequence. Since, our sample processing method enabled identification of all the species of ACB complex by PCR even in uncultured tracheal aspirates, adaptation of our protocol would enable same day (6 - 8 h) reporting and help the clinician make evidence based therapeutic decision quickly.展开更多
Brown rots of Red Fuji apples were observed in Hangzhou city (Zhengjiang Province, China). The causal agent was isolated and identified in both morphological and molecular genetic levels. The phenotype and phylogeneti...Brown rots of Red Fuji apples were observed in Hangzhou city (Zhengjiang Province, China). The causal agent was isolated and identified in both morphological and molecular genetic levels. The phenotype and phylogenetic analysis revealed that the isolate was </span><i><span style="font-family:Verdana;">Diaporthe phaseolorum</span></i><span style="font-family:Verdana;"> var. </span><i><span style="font-family:Verdana;">caulivora</span></i><span style="font-family:Verdana;">, and its pathogenicity on apple fruit was confirmed by re-inoculation experiment. To our knowledge, this is the first report of </span><i><span style="font-family:Verdana;">D. phaseolorum</span></i><span style="font-family:Verdana;"> var. </span><i><span style="font-family:Verdana;">caulivora</span></i><span style="font-family:Verdana;"> causing postharvest fruit rot on apple in China.展开更多
文摘目的建立荧光定量PCR检测美洲钩虫的方法。方法提取虫体基因组DNA、根据GenBank中美洲钩虫ITS-2序列设计特异引物,PCR扩增ITS-2序列、克隆、测序和比对。常规PCR检验引物特异性。将ITS-2序列扩增产物回收、纯化后经T克隆转入大肠埃希菌DH5α,提取重组质粒,鉴定后作为标准品模板建立荧光定量PCR标准曲线,并做灵敏性和重复性试验。结果构建的荧光定量PCR标准曲线循环阈值与模板浓度呈良好的线性关系,溶解曲线的吸收峰单一,实验重复性良好。结论成功构建了SYBR Green Ⅰ荧光定量PCR检测美洲钩虫的方法,可用于快速、准确、定量检测美洲钩虫。
文摘This paper provides a critical analysis of the design and implementation of Graduate English 1 for 1st year post graduates,9 credits to gain toward the degree.Specifically,the paper examines the details of the module specification template(MST) and provides an analysis on the aims,objectives,teaching methods,learning outcomes and assessment within the learning program,with reference to current theory on curriculum design and assessment.
文摘Currently, available phenotyping and commercial methods report <em>A. baumannii</em> only as <em>Acinetobacter calcoaceticus-baumannii</em> complex (ACB) and do not identify individual members of the complex. This is a single blind study aimed to evaluate certain commonly used species-specific genetic markers namely, Intergenic Transcribed Spacer region in 16S rRNA of <em>A. baumannii</em> (Ab-ITS) and <em>gyrB</em>, for identification of ACB members. These molecular targets were first validated on clinical isolates (n = 200) and subsequently on uncultured tracheal aspirates (n = 172). Among the clinical isolates, 183/200 (91.5%) were positive for Ab-ITS. The clinical isolates 17 (17/200) which are failed to amplify in Ab-ITS PCR were subsequently diagnosed by <em>gyrB</em> PCR as <em>A. calcoaceticus</em> (n = 2), <em>A. pitti</em> (n = 6) and <em>A. nosocomialis</em> (n = 9) but not <em>A. baumannii</em>. Among the tracheal aspirates, 62 samples were reported as sterile in Advanced Expert System of VITEK-2, among the remaining 110 samples, 68.1% (75/110) samples contained Ab-ITS target. Twenty-five of the sterile samples (25/62) were found to contain Ab-ITS target sequence. Since, our sample processing method enabled identification of all the species of ACB complex by PCR even in uncultured tracheal aspirates, adaptation of our protocol would enable same day (6 - 8 h) reporting and help the clinician make evidence based therapeutic decision quickly.
文摘Brown rots of Red Fuji apples were observed in Hangzhou city (Zhengjiang Province, China). The causal agent was isolated and identified in both morphological and molecular genetic levels. The phenotype and phylogenetic analysis revealed that the isolate was </span><i><span style="font-family:Verdana;">Diaporthe phaseolorum</span></i><span style="font-family:Verdana;"> var. </span><i><span style="font-family:Verdana;">caulivora</span></i><span style="font-family:Verdana;">, and its pathogenicity on apple fruit was confirmed by re-inoculation experiment. To our knowledge, this is the first report of </span><i><span style="font-family:Verdana;">D. phaseolorum</span></i><span style="font-family:Verdana;"> var. </span><i><span style="font-family:Verdana;">caulivora</span></i><span style="font-family:Verdana;"> causing postharvest fruit rot on apple in China.
