摘要
目的以猪附红细胞体(E.suis)为例,建立套式PCR方法,定性测定E.suis感染大鼠血液中DNA,检验套式PCR方法在其动物模型应用中的实用性和可操作性。方法用阳性血液样品分别经腹腔、皮下及肌肉感染大鼠,定期采血,进行常规血涂片染色,并从血液中分离纯化E.suis,提取DNA,进行套式PCR法扩增,凝胶电泳定性。结果套式PCR经扩增后得到了一长度为498bp的特异条带,测序鉴定确为目的片段;在整个实验过程中未见感染大鼠明显的临床症状。结论可以用大鼠建立猪附红细胞体动物感染动物模型;附红细胞体的感染方式有以下几种:腹腔、皮下及肌肉感染。在隐性感染的整个阶段,PCR方法依靠其高敏感性,作为猪附红细胞体感染大鼠模型检测指标之一,有其必要性和重要性。
Objective Two pairs of E. suis specific primers were designed to establish a new nested-PCR method in order to assess the E. suis DNA in the blood of E. suis infected rats. The practicability and operability of the nested-PCR techniques used in E.snis rat model were verified. Methods Rats were infected intraperitoneally, subcutaneously or intramuscularly with positive blood from infected swine. Blood samples were colleted and normal blood smear staining was done. The E. suis was purified and its DNA was extracted. Nested-PCR was done and the PCR products were assessed by electrophoresis in 0.85 % agarose gel. Results A 498 bp specific fragment was amplified with nested-PCR. It was verified by sequencing. During the experiment no obvious clinical symptom was found in infected rats. Conclusion A rat model of infection with eperythrozoon has been established. In the transcription latent period, nested-PCR is more sensitive and it may become an important and necessary technique in E. suis models assessment.
出处
《中国实验动物学报》
CAS
CSCD
2007年第2期142-145,共4页
Acta Laboratorium Animalis Scientia Sinica
基金
山东省医药卫生科技发展计划重点项目(编号:2005ZD05)
山东省自然科学基金项目(编号:Y2005C21)
关键词
支原体感染
聚合酶链反应
模型
动物
大鼠
Mycoplasma infections
Polymerase chain reaction
Models, animal
Rat
