炭疽与蜡样芽胞杆菌rpoB基因交叉反应的分子分析

发布时间：2018-01-12 13:17

本文关键词：炭疽与蜡样芽胞杆菌rpoB基因交叉反应的分子分析　出处：《现代预防医学》2017年12期 　论文类型：期刊论文

【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。
[Abstract]:Objective to find the PCR cross reaction between Bacillus anthracis and Bacillus cereus rpo B gene and study the cause of cross reaction. Methods Bacillus spp. Isolated from food and soil were stained with morphology. Biochemical reaction, time-of-flight mass spectrometry, protein toxin crystallization test, root growth test, anthrax phage lysis test, anthracis and Bacillus cereus fluorescence quantitative PCR identification. The rpo B gene real-time fluorescence quantitative PCR amplification products were cloned and sequenced, compared with the gene sequence. Some strains were typed by MLST. Results some Bacillus anthrax real-time fluorescence quantitative PCR test showed that rpo B amplification was positive, and it was identified as Bacillus cereus. RPO B amplification product was sequenced. Four new St types were found in different groups, which were similar to those of Bacillus anthracis. Conclusion there is PCR cross reaction between Bacillus cereus and rpo B gene of Bacillus anthracis. The molecular biological basis is the high similarity of the genomic sequence, which has little relationship with the genetic variation of the strain. The genetic diagnosis of Bacillus anthracis and Bacillus cereus should refer to the joint identification of toxin plasmid and chromosome gene.
【作者单位】：陕西省疾病预防控制中心;浙江中医药大学;
【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)
【分类号】：R440
【正文快照】： 蜡样芽胞菌群是指一群在基因组水平上具有高度相似性的细菌,包括蜡样芽胞杆菌、炭疽芽胞杆菌、苏云金杆菌等6种芽胞杆菌[1]。其中炭疽芽胞杆菌会产生致死毒素和水肿毒素,引起人和动物的烈性传染病炭疽。蜡样芽胞杆菌在自然界分布广泛,为一种条件致病菌,产生肠毒素,引起食物中

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