炭疽与蜡样芽胞杆菌rpoB基因交叉反应的分子分析  

Molecular analysis on cross-reaction of rpoB gene between Bacillus anthracis and Bacillus cereus

作　　者：李文涓[1] 石一[1] 李沈玲[2] 王维华[1] 张铮[1] 马琳[1] 刘长宏[1] 马国柱[1] 刘东立[1] 

机构地区：[1]陕西省疾病预防控制中心,陕西西安710054 [2]浙江中医药大学,浙江杭州310053

出　　处：《现代预防医学》2017年第12期2238-2244,共7页Modern Preventive Medicine

基　　金：陕西省科技资源开放共享平台项目(2016FWPT-12);陕西省卫生计生科研基金(2016D094)

摘　　要：目的发现炭疽芽胞杆菌与蜡样芽胞杆菌rpo B基因PCR交叉反应,对产生交叉反应的原因进行研究。方法对分离自食品及土壤的芽胞杆菌进行形态染色、生化反应、飞行时间质谱、蛋白毒素结晶试验、根状生长试验、炭疽噬菌体裂解试验、炭疽杆菌及蜡样杆菌荧光定量PCR等鉴定,并对rpo B基因实时荧光定量PCR扩增产物进行克隆测序,比对基因序列。对部分菌株进行MLST分型,探讨交叉反应的遗传关系。结果部分芽胞杆菌炭疽实时荧光定量PCR试验出现rpo B扩增阳性,经鉴定为蜡样芽胞杆菌。rpo B扩增产物测序显示,与炭疽杆菌高度相似。MLST分型显示分布在不同的群,发现4种新的ST型。结论蜡样芽胞杆菌与炭疽芽胞杆菌的rpo B基因存在PCR交叉反应,其分子生物学基础是基因组序列的高度相似性,与菌株遗传变异关系不大,对炭疽杆菌及蜡样杆菌的基因诊断应该参考毒素质粒及染色体基因的联合鉴定。The rpoB gene PCR cross-reactivity between Bacillus anthrax and Bacillus cereus in Shanxi of China wasreported.The Bacillus isolated from food and soil was identifiedbased on morphological, biochemical reaction, MOLDI-TOF,protein toxin crystal test, root growth test, anthracis phage lysis test, Bacillus anthraci and Bacillus cereus real-time PCR.The rpoB gene real-time PCR amplification, cloning and sequencing, analyzing and alignment of the gene sequence wereconducted. The results indicated that some of Bacillus real-time PCR showed Bacillus athracis rpoB gene was positive andidentified as Bacillus cereus. PCR amplification product sequencing showed that the high similarity with Bacillus anthricais.So the main reason of rpoB gene PCR cross-reaction between Bacillus cereus and Bacillus anthracis was highly similargenome sequences. Muhilocus sequence typing （MIST） of 9 Bacillus ceres strains showed in different groups, and four newST profiles were found. PCR cross-reaction between strains had little genetic variation. Gene diagnosis of Bacillus anthracisand Bacillus cereus should be combined with identification of toxin plasmid gene and chromosome gene.

关 键 词：炭疽芽胞杆菌 蜡样芽胞杆菌 PCR交叉反应 多位点序列分子分型(MLST) 

分 类 号：R378[医药卫生—病原生物学]