大鼠肺炎支原体的分离鉴定及致病性研究OA
Isolation,identification and pathogenicity of Mycoplasma pneumoniae in rats
目的 从某发生呼吸道病的大鼠养殖场分离肺炎支原体(Mycoplasma pneumoniae,MP)并鉴定该病原,进行大鼠致病性实验,以期获得目前流行病病原体.方法 对疑似MP感染大鼠肺组织PCR检测,扩增得到约为280 bp的单一条带.病料接种支原体肉汤培养基,并在支原体琼脂培养基上纯化.通过菌落特征、生化特性、16SrRNA扩增和同源性分析对该菌进行鉴定.分离菌株大鼠致病性实验:用浓度为1 x 107 CCU/mL菌液对5周龄SD大鼠滴鼻,观察其临床症状和病理变化.结果 通过PCR方法初步确定疑似病例为大鼠肺炎支原体感染,接种肺支原体肉汤培养基生长情况符合该菌培养特点,纯化后菌落呈圆形,经Giemsa染色后呈淡紫色;生化结果为分解葡萄糖产酸、不水解精氨酸和尿素;16S rRNA扩增和测序结果与鼠源MP菌株(AL445565.1)同源性相似度高达99.05%,表明分离获得大鼠肺炎支原体.该病原攻毒SD大鼠后出现呼吸道症状,肺部有出血、肉变,组织有大量炎性细胞浸润且MP PCR鉴定结果呈阳性.结论 本实验成功分离了一株大鼠MP,该株MP能致大鼠发生典型肺炎症状,可为进一步研究大鼠支原体肺炎的防控奠定良好基础.
Objective Mycoplasma pneumoniae(MP)was isolated from a rat farm with respiratory diseases and the pathogen was identified.A rat pathogenicity experiment was conducted to obtain the current prevalent pathogen.Methods Lung tissues were isolated from rats with suspected MP infection and analyzed by PCR,yielding an amplified single band of approximately 280 bp.The samples were inoculated into mycoplasma broth medium and further purified on mycoplasma agar plates.The isolate was identified based on colony morphology,biochemical characteristics,amplification of the 16S rRNA gene,and homology analysis.Pathogenicity tests were conducted in 5-week-old SD rats by intranasal inoculation with a bacterial suspension at a concentration of 1 × 107 CCU/mL,and the subsequent clinical symptoms and pathological changes were observed.Results PCR confirmed that the suspected cases were infected with MP.The growth characteristics in mycoplasma broth medium matched those of the target organism,and the purified colonies appeared round and stained light purple with Giemsa stain.Biochemical tests revealed acid production from glucose fermentation but no hydrolysis of arginine or urea.Amplification and sequencing of the 16S rRNA gene showed high homology(99.05%)with the rat-derived MP strain(AL445565.1),confirming successful isolation of MP.Rats challenged with the isolate developed respiratory symptoms,including pulmonary hemorrhage,consolidation,massive infiltration of inflammatory cells,and positive MP PCR result in lung tissues.Conclusions This study successfully isolated a strain of MP from rats,which induced typical pneumonia symptoms in SD rats.These findings provide a solid foundation for further research into the prevention and control of MP in rats.
刘骁霖;王晓斌;刘硕;曹卫强;任守海;于永乐;杨瑞梅
青岛农业大学动物医学院,山东青岛 266109青岛农业大学动物医学院,山东青岛 266109青岛农业大学动物医学院,山东青岛 266109青岛大任富城畜牧有限公司,山东青岛 266109青岛大任富城畜牧有限公司,山东青岛 266109青岛农业大学动物医学院,山东青岛 266109青岛农业大学动物医学院,山东青岛 266109
生物科学
大鼠肺炎支原体PCR分离鉴定致病性
ratsMycoplasma pneumoniaePCRisolationidentificationpathogenicity
《中国实验动物学报》 2026 (2)
228-237,10
白鼠疫病防控技术服务协议(横20230227).Funded by the Technical Service Agreement for White Rat Disease Prevention and Control(Heng 20230227).
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