中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 美国白蛾核型多角体病毒实时荧光定量PCR检测方法的建立及应用 昆虫学报 2010,53 (7) 106-112
Position: Home > Articles > Establishment and application of real-time fluorescence quantitative PCR for detection of Hyphantria cunea nucleopolyhedrovirus Acta Entomologica Sinica 2010,53 (7) 106-112

美国白蛾核型多角体病毒实时荧光定量PCR检测方法的建立及应用

作  者：
乔鲁芹;曲良建;王玉珠;张永安;杨忠岐;陶万强;关玲
单  位：
北京市林业保护站;中国林业科学研究院森林生态环境与保护研究所;山东农业大学植物保护学院
关键词：
美国白蛾核型多角体病毒;SYBR GreenⅠ;荧光定量PCR;感病幼虫;检测
摘  要：
为了建立对昆虫核型多角体病毒(nucleopolyhedrovirus)进行定量检测方法,本研究选用美国白蛾Hyphantria cunea核型多角体病毒的polyhedrin基因为目的基因,经引物设计、PCR扩增、目的片段与载体连接转化以及重组质粒的鉴定,并对重组质粒标准品和样品DNA进行检测,构建出美国白蛾核型多角体病毒SYBR GreenⅠ荧光定量标准曲线。统计学分析显示标准品浓度的对数值与Ct值之间存在良好的线性关系(R=0.998)。该方法的检测灵敏度为101～102拷贝/μL,线性范围达5个数量级,扩增产物形成单一的特异性熔解峰,组内组间的变异系数均小于3%。根据已建立的方法对不同感染时间段的感病幼虫进行检测,每毫克幼虫样本内病毒polyhedrin基因拷贝数增殖倍数对数值与感染时间(d)呈正相关(R=0.987)。这些结果表明,建立的美国白蛾核型多角体病毒荧光定量PCR检测方法是可靠的,具有灵敏度高、重复性好等特点,可为昆虫核型多角体病毒体内增殖动态研究及昆虫病毒的标准化生产提供参考。
译  名：
Establishment and application of real-time fluorescence quantitative PCR for detection of Hyphantria cunea nucleopolyhedrovirus
作  者：
QIAO Lu-Qin2, QU Liang-Jian1, WANG Yu-Zhu1, ZHANG Yong-An1,, YANG Zhong-Qi1, TAO Wan-Qiang3, GUAN Ling3 (1. Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry, Beijing 100091, China; 2. College of Plant Protection, Shandong Agricultural University, Tai'an, Shandong 271018, China; 3. Forestry Protection Center in Beijing City, Beijing 100029, China)
关键词：
Hyphantria cunea nucleopolyhedrovirus; SYBR Green Ⅰ; fluorescence quantitative PCR; infected larvae; detection
摘  要：
Primers were designed based on polyhedrin from Hyphantria cunea nucleopolyhedrovirus in order to establish the detection method of SYBR GreenⅠ real-time fluorescence quantitative PCR for HcNPV, and the standard curve of SYBR GreenⅠreal-time fluorescence quantitative PCR for polyhedrin was established through PCR amplification, ligation of target genes and vector, transformation, identification of recombinant plasmid and DNA sample detection. Statistic analysis showed that there was a good linear relationship between Ct value and the logarithmic value of plasmid concentrations (R=0.998). The sensitivity of the method was 101-1×102 copies/μL, and wide detection range of 5 orders of magnitude was obtained. Larvae infected at different time were sampled and detected by the method, and the results showed that there was a good linear relation between the logarithmic value of the multiples of gene copies and the infection time (R= 0.987). The results show that the detection method for HcNPV is of high sensitivity, reproducibility and credibility, which will facilitate the epizootiology and the standardized production for insect virus.

相似文章

导出引用
计量
文章访问数: 7
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46