胸膜肺炎放线杆菌无耐药标记的活的、减毒的apxIICA失活突变株的构建与鉴定

Construction and characterization of a live, attenuated apxIICA inactivation mutant of Actinobacillus pleuropneumoniae lacking a drug resistance marker.

作者信息

Bei Weicheng, He Qigai, Yan Lin, Fang Liurong, Tan Yadi, Xiao Shaobo, Zhou Rui, Jin Meilin, Guo Aizhen, Lv Jianqiang, Huang Hongliang, Chen Huanchun

机构信息

Laboratory of Animal Infectious Diseases, State Key Laboratory of Agricultural Microbiology, College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei 430070, China.

出版信息

FEMS Microbiol Lett. 2005 Feb 1;243(1):21-7. doi: 10.1016/j.femsle.2004.11.033.

DOI:10.1016/j.femsle.2004.11.033

PMID:15667996

Abstract

The apxIIC gene of Actinobacillus pleuropneumoniae serotype 7 was inactivated by homologous recombination using a sucrose counter-selectable marker system, resulting in a mutant strain that had no antibiotic resistance marker and expressed an inactivated ApxII toxin. The safety and immunogenicity of the mutant were evaluated in mice. The mutant strain caused no adverse effects in mice at doses up to 2 x 10(9) CFU via the intraperitoneal route while the parental strain induced total mortality at a dose of 2 x 10(7) CFU. Mice vaccinated intraperitoneally with the mutant strain had 100% and 70% protection against homologous (serotype 7) or heterologous (serotype 1, 3) challenge with A. pleuropneumoniae, respectively. The A. pleuropneumoniae mutant strain HB04C- and the counterselection method used in the study show promise in developing effective live vaccines for porcine pleuropneumonia and for other infections diseases of the respiratory system.

摘要

利用蔗糖反选择标记系统通过同源重组使胸膜肺炎放线杆菌血清型7的apxIIC基因失活，从而获得了一个无抗生素抗性标记且表达失活ApxII毒素的突变菌株。对该突变株在小鼠体内的安全性和免疫原性进行了评估。该突变株经腹腔途径给予高达2×10⁹CFU的剂量时，对小鼠未产生不良影响，而亲本菌株在2×10⁷CFU的剂量下可导致全部死亡。经腹腔接种该突变株的小鼠分别对同源（血清型7）或异源（血清型1、3）胸膜肺炎放线杆菌攻击具有100%和70%的保护作用。本研究中所用的胸膜肺炎放线杆菌突变株HB04C-及反选择方法在开发用于猪胸膜肺炎及其他呼吸系统感染疾病的有效活疫苗方面显示出前景。