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。
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. coll. The optimized Mg^2＋ concentration of this method is 5 mmool/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 eontroled 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 eontroled in esophagus and duodenum extreme dramatically; And it is a little higher than the controled in jejunum, ileum and cecum, and lower than the eontroled a little in rectum.
出处
《四川农业大学学报》
CSCD
2006年第3期331-336,共6页
Journal of Sichuan Agricultural University
基金
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)
