Genetic Characterization of Clostridium botulinum Associated with Type B Infant Botulism in Japan
Genetic Characterization of Clostridium botulinum Associated with Type B Infant Botulism in Japan
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DOI：
10.1128/jcm.00077-09
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发表时间：
2009-09-01
影响因子：
9.4
通讯作者：
Kozaki, Shunji
中科院分区：
文献类型：
--
作者：
Umeda, Kaoru;Seto, Yoshiyuki;Kozaki, Shunji
采用boNT/ B基因序列系统发育分析、基因分型及脉冲场凝胶电泳（PFGE）测定boNT/ B基因定位进行日本婴儿肉毒中毒分子流行病学分析，对15株B型肉毒梭菌（其中3株与日本婴儿肉毒中毒相关）进行遗传特征鉴定。菌株Osaka05从2005年1例病例中分离得到，具有独特的boNT/ B基因序列，经系统发育分析认为是boNT/ B新亚型。2006年从一个病例中分离出的菌株Osaka06被归类为B2亚型，与1995年从一个病例中分离出的菌株111相同。在美国与婴儿肉毒杆菌中毒相关的五个分离株被归类为B1亚型。日本食品样品分离株分为B1和B2亚型，但PFGE基因分型未显示与婴儿肉毒杆菌中毒有关。与未消化DNA的PFGE和Southern blot杂交结果表明，boNT/ B基因位于来自不同来源的3种boNT/ B亚型的5个菌株的大质粒（约150 kbp， 260 kbp， 275 kbp或280 kbp）上。利用重组BoNT/ b - c末端结构域的夹心酶联免疫吸附实验表明，Osaka05的肉毒杆菌神经毒素（BoNT）具有不同于BoNT/B1和BoNT/B2的抗原性。我们建立了BoNT/B亚型多重PCR检测方法，这将为B型菌株及其引起的传染病的流行病学研究提供依据。
The 15 proteolytic Clostridium botulinum type B strains, including 3 isolates associated with infant botulism in Japan, were genetically characterized by phylogenetic analysis of boNT/ B gene sequences, genotyping, and determination of the boNT/ B gene location by using pulsed-field gel electrophoresis (PFGE) for molecular epidemiological analysis of infant botulism in Japan. Strain Osaka05, isolated from a case in 2005, showed a unique boNT/ B gene sequence and was considered to be a new BoNT/B subtype by phylogenetic analysis. Strain Osaka06, isolated from a case in 2006, was classified as the B2 subtype, the same as strain 111, isolated from a case in 1995. The five isolates associated with infant botulism in the United States were classified into the B1 subtype. Isolates from food samples in Japan were divided into the B1 and the B2 subtypes, although no relation with infant botulism was shown by PFGE genotyping. The results of PFGE and Southern blot hybridization with undigested DNA suggested that the boNT/ B gene is located on large plasmids (approximately 150 kbp, 260 kbp, 275 kbp, or 280 kbp) in five strains belonging to three BoNT/B subtypes from various sources. The botulinum neurotoxin (BoNT) of Osaka05 was suggested to have an antigenicity different from the antigenicities of BoNT/B1 and BoNT/B2 by a sandwich enzyme-linked immunosorbent assay with the recombinant BoNT/B-C-terminal domain. We established a multiplex PCR assay for BoNT/B subtyping which will be useful for epidemiological studies of type B strains and the infectious diseases that they cause.