炭疽芽孢杆菌的分离鉴定及其与疫苗株的鉴别诊断  

Isolation and identification of Bacillus anthracis strains and the differential diagnosis of vaccine strains

作　　者：祝令伟[1] 郭学军[1] 李吉平[1] 顾贵波 刘军[1] 纪雪[1] 冯书章[1] 孙洋[1] 

机构地区：[1]军事医学科学院军事兽医研究所,吉林省人兽共患病预防与控制重点实验室,长春130122 [2]辽宁省动物疫病预防与控制中心,沈阳110164

出　　处：《中国人兽共患病学报》2016年第11期983-986,共4页Chinese Journal of Zoonoses

基　　金：吉林省青年科研基金(No.20150520130JH);农业部《动物疫情监测与防治(兽医)项目》(No.271)联合资助~~

摘　　要：目的建立炭疽芽孢杆菌临床分离株和疫苗株的鉴别诊断方法,为炭疽疫情判断和疾病控制提供依据。方法以炭疽芽孢外壁胶原样蛋白的编码基因bclA和pXO1质粒上的aat序列为靶点,设计引物,通过PCR扩增片段多态性和测序结果分析可变串联重复序列(VNTR)。在生物安全3级实验室分离炭疽菌体蛋白,使用MALDL-TOF/TOF质谱仪进行检测,BioTyper软件分析图谱。结果炭疽分离株与Ⅱ号、无荚膜疫苗株,VNTR分析结果均有不同。2个分离株测定bclA基因序列全长均为1 113bp,编码的蛋白包含66个GXX重复和5个BclA重复。质谱分析发现,2个分离株菌体蛋白表达峰图基本一致,而2个疫苗株与之相比有很大的差异,许多蛋白的表达明显下调。结论两种方法都能够有效鉴别炭疽临床分离株和疫苗株。The differential diagnosis methods were developed to identify the Bacillus anthracis clinical strains and vaccine strains.Firstly,the variable-number tandem repeat（VNTR）analysis method was created basing on the bclAgene of exosporium collagen-like protein.It was founded that the PCR amplification fragment length of the two isolated strains was different from the II vaccine strain and non-capsulated vaccine strain.The BclA protein of isolated strain contain 66 copies of GXX repeat and five copies of（GPT）5GDTGTT（BclA repeat）.Secondly,the bacterial protein expression level of isolated strain was also different from the vaccine strains by MALDI-TOF/TOF mass spectrometry analysis.Both of the two methods can effectively identify the Bacillus anthracis clinical isolated strains and vaccine strains.

关 键 词：炭疽芽孢杆菌 疫苗株 鉴别诊断 质谱 可变串联重复序列 

分 类 号：R378.7[医药卫生—病原生物学]