四川农业大学学报 ›› 2006, Vol. 24 ›› Issue (03): 331-336.doi: 10.16036/j.issn.1000-2650.2006.03.019

• 研究论文 • 上一篇    下一篇

16SrDNA实时荧光定量PCR检测DPV强毒感染鸭气管及消化道大肠杆菌动力学

胡骑1,2, 程安春1,2, 汪铭书1,2, 刘艳丽1,2, 葛忠源1,2, 黄永成1,2, 张素辉1,2, 杨晓燕1,2, 齐雪峰1,2, 陈孝跃1,2   

  1. 1. 四川农业大学 动物科技学院, 四川 雅安 625014;
    2. 动物疫病与人类健康四川省重点实验室, 四川 雅安 625014
  • 收稿日期:2006-04-24 出版日期:2006-09-30 发布日期:2017-03-06
  • 基金资助:
    国家科技攻关重大项目(2004BA901A03);教育部"新世纪优秀人才支持计划"项目(NCET-04-0906);四川省重点建设学科项目(SZD0418)

16SrDNA Real-time Fluorescence Quantitative PCR to Detect the Dynamics of Escherichia Genus in Trachea and Alimentary Canal of Ducks Infected by DPV Virulent Strain Artificially

HU Ji1,2, CHENG An-chun1,2, WANG Ming-shu1,2, LIU Yan-li1,2, GE Zhong-yuan1,2, HUANG Yong-cheng1,2, ZHANG Su-hui1,2, YANG Xiao-yan1,2, QI Xue-fen1,2, CHEN Xiao-yue1,2   

  1. 1. College of Animal and Technology, Sichuan Agricultural University, Yaan 625014, Sichuan, China;
    2. The Key Laboratory of Animal Disease and Human Health of Sichuan Province, Yaan 625014, Sichuan, China
  • Received:2006-04-24 Online:2006-09-30 Published:2017-03-06

摘要: 参照Genebank大肠埃希氏菌属(E. coli)16SrDNA保守基因序列设计并合成定量PCR引物及针对大肠杆菌属的Taqman探针,以E. coli标准菌株的PCR扩增产物作为阳性模板制作标准曲线,建立检测E. coli的定量PCR方法。该法Mg2+最佳工作浓度5 mmol/L,能够定量和特异检测E. coli而对肠道优势菌群的代表种葡萄球菌、双歧杆菌呈阴性。检测E. coli的灵敏度可达3个/mL。利用该法对DPV强毒感染鸭急性病例的气管和消化道E. coli检测,结果表明:气管E. coli数量普遍低于对照。食道波动巨大,普遍高于对照;十二指肠、空肠变化不明显;回肠波动巨大,总体高于对照;盲肠显著低于对照;直肠与对照差异不显著。DPV感染致死鸭的气管E. coli数量显著低于对照;食道和十二指肠极显著高于对照;空肠、回肠和盲肠均略高于对照;直肠略低于对照。

关键词: 实时荧光定量PCR, DPV强毒感染鸭, 气管, 消化道, 大肠杆菌, 检测

Abstract: One pair of quantitative PCR primer and one specific Escherichia genus(E. coli) Taqman probe are designed and synthesized by consulting the Escherichia Genus 16SrDNA conservative gene sequence in Genbank. The PCR amplification products of E. coli standard strain are used as masculine templates to make the standard curve for founding the quantitative PCR method to detect E. coli. The optimized Mg2+ concentration of this method is 5 mmol/L, and this method can be used for detecting E. coli quantitatively and specifically. But other predominant intestinal bacteria such as Staphylococcus and Bifidobacterium can not be detected. The sensation of it can be 3/mL. The results demonstrate that:the number of E. coli in trachea is lower than the controled in general. It waves dramatically in esophagus, and higher than the controled in general. It is not waved obviously in duodenum and jejunum, but dramatically in ileum, higher than the controled in general. It is lower than the controled dramatically in cecum, and not obviously in rectum. The number of E. coli of dead ducks infected by DPV in trachea is lower than the controled dramatically, and higher than the controled in esophagus and duodenum extreme dramatically;And it is a little higher than the controled in jejunum, ileum and cecum, and lower than the controled a little in rectum.

Key words: real-time FQ-PCR, DPV virulent strain infected ducks, trachea, alimentary tract, E. coli, detection

中图分类号: 

  • S858.32