小附红细胞体的分离鉴定及其对猪的人工感染试验  

Isolation and identification of Mycoplasma parvum and its artificial infection experiment in swine

作　　者：雷晓思 周鹏[2,3] 米荣升[2,3] 黄燕[2,3] 刘宇轩[2,3] 胡盼[2,3] 詹婷婷[2] 江帆[1] 夏延富 邓俊良[1] 陈兆国[2,3] 

机构地区：[1]四川农业大学动物医学院动物疫病与人类健康四川省重点实验室,四川雅安625014 [2]中国农业科学院上海兽医研究所农业部动物寄生虫学重点实验室农业部动物产品质量安全生物性危害因子风险评估实验室(上海),上海200241 [3]江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009 [4]惠州市惠阳区动物疫病预防控制中心,广东惠州516211

出　　处：《中国兽医科学》2015年第2期142-149,共8页Chinese Veterinary Science

基　　金：教育部"长江学者和创新团队发展计划"创新团队项目(IRT0848);公益性行业(农业)科研专项(200903036);中央级公益性科研院所基本科研业务费专项(2014JB04)

摘　　要：采用2对附红细胞体种特异性引物,对上海地区2个屠宰场采集的736份猪血液基因组DNA进行PCR检测,将小附红细胞体特异性引物扩增产物克隆到pMD18-T载体,进行序列测定和生物信息学分析。将小附红细胞体阳性血液分离物肌肉注射给5头切除脾的断奶广西巴马小型猪仔猪,感染后定期采血,进行显微镜检,并提取基因组DNA进行PCR检测。另设3头猪作为不感染对照组,进行相同的检测。结果显示,共有101份样品检测出附红细胞体阳性,阳性率为13.72%,其中54份为小附红细胞体单独感染,占7.34%。5头巴马小型猪在接种小附红细胞体后第13~16天起,至第34天试验结束,出现明显的临床症状;血液显微镜检和PCR检测均发现存在小附红细胞体感染;其序列测定显示,16SrRNA基因的部分序列与接种的小附红细胞体的相应序列完全一致。上述结果表明,本研究成功筛选出小附红细胞体阳性血液,并成功实现了小附红细胞体对巴马小型猪的人工感染试验。A total of 736 swine blood samples were collected from two slaughterhouses in Shanghai,and their genomic DNAs were extracted and amplified by PCR with two pairs of Mycoplasmaspecies specific primers.The PCR products were cloned into pMD18-T vector,sequenced and analyzed with the bioinformatics software.The isolates of M.parvuminfective blood samples were inoculated intramuscularly to 5splenectomized weaning Guangxi Bama miniature pigs.The blood samples of the Bama miniature pigs were collected periodically after infection and detected by microscopy and the PCR.Other 3pigs,which were not infected as the negative control,were also detected.In result,101 samples were identified as Mycoplasma spp.positive and the infection rate was 13.72%,of which 54 samples were M.parvuminfection alone,account for 7.34%.From day 13 or 16post-infection to day 34post-infection,5Bama miniature pigs appeared obvious clinical symptoms.All the blood samples were M.parvuminfection by microscopy and PCR detection.The partial 16 SrRNA sequences amplified by the specific PCR showed that they were belong to M.parvumsequence in accordance with that of the inoculation samples.The above-mentioned results showed that the positive porcine blood samples with M.parvum were successfully selected and identified,and the artificial infection experiment was successfully established in this study.

关 键 词：小附红细胞体 分离 鉴定 猪 人工感染 

分 类 号：S858.28[农业科学—临床兽医学]