凹甲陆龟沙门氏菌的分离鉴定及毒力因子基因的PCR检测  

Isolation and Identification of Salmonella in Manouria impressa and Detection of Virulence Factor Gene by PCR

作　　者：黄道超[1,2] 姜波[2] 杨光友[2,3] 牛李丽[3] 王强[3] 余星明[3] 邓家波[3] 赵波[3] 陈维刚[3] 

机构地区：[1]重庆医科大学附属儿童医院动物实验中心,重庆400014 [2]四川农业大学动物疫病与人类健康四川省重点实验室,雅安625014 [3]成都动物园,成都610041

出　　处：《生物技术通报》2009年第10期156-160,共5页Biotechnology Bulletin

基　　金：成都大熊猫繁育研究基金会项目(CPF008);重庆医科大学校级课题项目(XBYB2008043)

摘　　要：从成都动物园因腹泻死亡的凹甲陆龟体内分离到一株病源菌,经选择性分离培养、生化试验、血清型鉴定等,确定该病源菌为D群肠炎沙门氏菌,其抗原结构式为1,9,12∶g,m∶-。该菌株对小鼠有较强的致病性,能引起人工感染小鼠大量死亡,且从其体内分离到相同特性的菌株;药物敏感性试验证实该分离菌对头孢曲松、氧氟沙星、卡拉霉素、多粘菌素、复方新诺明、呋喃唑酮等敏感,对链霉素、四环素、氯霉素等耐药;同时根据GenBank中已报道的沙门氏菌毒力因子ivnA和ivnE的基因序列,设计两对特异性引物对该菌进行PCR检测,结果在该菌中成功扩增并检测出ivnA和ivnE基因。本试验尝试了沙门氏菌的PCR检测方法,为该分子诊断技术的全面推广奠定了良好基础,也为今后该疾病的预防、诊断和治疗提供了科学的实验数据。A bacteria strain of Salmonella was isolated from a dead Manouria impressa suffered with diarrhea in Chengdu-Zoo. After culture, isolation, biochemical reaction and serological test, it was identified as Salmonella enteritidis of group D, and its antigenic structural formula was 1,9,12：g, m： -. The strain with high pathogenicity caused mass mortality of mouses infected artificially, and the same strain was isolated from dead mouses. In drug sensitivity test, the strain was sensitive for ceftriaxone, Ofloxacin, kalamycin, polymyxin, SMZ-TMP, furaltadone, and resistant for phytomycin, ambramycin, amphemycin. Meanwhile, according to the sequences of ivnA and ivnE genes reported in GenBank, two pairs of specific primers were designed for detection of virulence factor genes. At resuits, ivnA and ivnE genes were amplified and detected successfully by PCR. The study provided a basis for the universal use of molecular diagnostic technique, and supplied scientific experimental datas for preventing, diagnosing and curing the disease in future.

关 键 词：凹甲陆龟 沙门氏菌 分离 鉴定 毒力因子基因PCR 

分 类 号：S947[农业科学—水产养殖]