[1]赵碧,熊朝丽,徐茂文,等.鸭肠炎病毒核衣壳蛋白质互作蛋白质基因荧光定量PCR检测方法的建立[J].江苏农业学报,2015,(02):389-393.[doi:10.3969/j.issn.1000-4440.2015.02.026]
 ZHAO Bi,XIONG Chao-li,XU Mao-wen,et al.Establishment of a real-time fluorescent quantitative PCR method for detecting the interacting protein (PKCI)gene of nucleocapsid protein of duck enteritis virus[J].,2015,(02):389-393.[doi:10.3969/j.issn.1000-4440.2015.02.026]
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鸭肠炎病毒核衣壳蛋白质互作蛋白质基因荧光定量PCR检测方法的建立()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年02期
页码:
389-393
栏目:
畜牧兽医·水产养殖
出版日期:
2015-04-30

文章信息/Info

Title:
Establishment of a real-time fluorescent quantitative PCR method for detecting the interacting protein (PKCI)gene of nucleocapsid protein of duck enteritis virus
作者:
赵碧1熊朝丽1徐茂文1周碧君12程振涛12文明12
(1.贵州大学动物科学学院,贵州贵阳550025;2.贵州省动物疫病与兽医公共卫生重点实验室,贵州贵阳550025)
Author(s):
ZHAO Bi1XIONG Chao-li1XU Mao-wen1ZHOU Bi-jun12CHENG Zhen-tao12WEN Ming12
(1.College of Animal Science, Guizhou University, Guiyang 550025,China;2.Key Laboratory of Animal Disease and Veterinary Public Health of Guizhou Province, Guiyang 550025,China)
关键词:
鸭肠炎病毒核衣壳蛋白质蛋白质激酶C抑制蛋白质荧光定量PCR
Keywords:
duck enteritis virusnucleocapsid proteinprotein kinase C inhibitor(PKCI)real-time fluorescent quantitative PCR
分类号:
S858.32
DOI:
10.3969/j.issn.1000-4440.2015.02.026
文献标志码:
A
摘要:
为进一步阐明鸭肠炎病毒(DEV)的致病机理,本试验建立检测鸭肠炎病毒核衣壳蛋白质(NP)的互作蛋白质(PKCI)基因荧光定量PCR方法。针对PKCI基因设计特异性引物,经PCR扩增目的基因构建重组质粒pMD18-T-PKCI,将其作为阳性标准品构建荧光定量PCR标准曲线,并对建立的荧光定量PCR方法进行重复性、特异性和敏感性试验。结果显示:标准曲线的线性关系为Y=-3.31x+42.00,相关系数为-1.00,扩增效率为100%,熔解曲线仅出现单特异峰,对H5亚型、H7亚型和H9亚型禽流感病毒及新城疫病毒、鸭肝炎病毒均未检测到荧光信号。表明本试验建立的方法具有良好的稳定性、特异性和灵敏性。
Abstract:
To identify the pathogenesis of duck enteritis virus(DEV), a real-time fluorescent quantitative PCR assay for detecting the interacting protein (PKCI) gene of nucleocapsid protein(NP) of duck enteritis virus was developed. With designed specific primer, the PKCI gene was amplified by PCR. The recombinanted plasmid of pMD18-T-PKCI was used as positive standard to construct a standard curve by real-time fluorescent quantitative PCR, Y=-3.31 x +42.00, with the correlation coefficient of -1.00 and the amplification efficiency of 100%. Melting curve appeared a single specific peak. The fluorescence signal were not detected in H5,H7,H9 subtype avian influenza viruses, newcastle disease virus, and duck hepatitis virus. The real-time fluorescent quantitative PCR method established in this study exhibited good stability, specificity and sensitivity.

参考文献/References:


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备注/Memo

备注/Memo:
收稿日期:2014-08-12 基金项目:国家自然科学基金项目(31260607);贵州省重点实验室培育项目[黔科合计Z字(2011)4008号] 作者简介:赵碧(1991-),男,云南曲靖人,硕士研究生,主要从事兽医微生物学与免疫学研究。(Tel)0851-8298078;(E-mail)710215448@qq.com 通讯作者:文明,(E-mail)as.mwen@gzu.edu.cn
更新日期/Last Update: 2015-04-30