Department of Medical Sciences, National Institute of Health, Nonthaburi, Thailand.
Department of Veterinary Science, Osaka Prefecture University, Osaka, Japan.
PLoS One. 2014 Jan 27;9(1):e77792. doi: 10.1371/journal.pone.0077792. eCollection 2014.
Thailand has had several foodborne outbreaks of botulism, one of the biggest being in 2006 when laboratory investigations identified the etiologic agent as Clostridium botulinum type A. Identification of the etiologic agent from outbreak samples is laborious using conventional microbiological methods and the neurotoxin mouse bioassay. Advances in molecular techniques have added enormous information regarding the etiology of outbreaks and characterization of isolates. We applied these methods in three outbreaks of botulism in Thailand in 2010.
METHODOLOGY/PRINCIPAL FINDINGS: A total of 19 cases were involved (seven each in Lampang and Saraburi and five in Maehongson provinces). The first outbreak in Lampang province in April 2010 was associated with C. botulinum type F, which was detected by conventional methods. Outbreaks in Saraburi and Maehongson provinces occurred in May and December were due to C. botulinum type A1(B) and B that were identified by conventional methods and molecular techniques, respectively. The result of phylogenetic sequence analysis showed that C. botulinum type A1(B) strain Saraburi 2010 was close to strain Iwate 2007. Molecular analysis of the third outbreak in Maehongson province showed C. botulinum type B8, which was different from B1-B7 subtype. The nontoxic component genes of strain Maehongson 2010 revealed that ha33, ha17 and botR genes were close to strain Okra (B1) while ha70 and ntnh genes were close to strain 111 (B2).
CONCLUSION/SIGNIFICANCE: This study demonstrates the utility of molecular genotyping of C. botulinum and how it contributes to our understanding the epidemiology and variation of boNT gene. Thus, the recent botulism outbreaks in Thailand were induced by various C. botulinum types.
泰国曾发生过几起因食用受污染食物而导致的肉毒中毒事件,其中最大的一起发生在 2006 年,当时实验室调查确定病原体为 A 型肉毒梭菌。使用传统的微生物学方法和神经毒素小鼠生物测定法从暴发样本中鉴定病原体非常繁琐。分子技术的进步为暴发的病因和分离株的特征提供了大量信息。我们在 2010 年泰国的三起肉毒中毒暴发中应用了这些方法。
方法/主要发现:共涉及 19 例病例(分别在廊开府、沙拉武里府各 7 例,在黎逸府 5 例)。2010 年 4 月在廊开府发生的首例暴发与 F 型肉毒梭菌有关,该菌是通过传统方法检测到的。沙拉武里府和黎逸府的暴发分别发生在 5 月和 12 月,是由 A1(B)型和 B 型肉毒梭菌引起的,这两种菌是通过传统方法和分子技术分别鉴定的。系统发育序列分析结果表明,沙拉武里 2010 年 A1(B)型菌株与岩手 2007 年菌株接近。对黎逸府第三次暴发的分子分析显示为 B8 型肉毒梭菌,与 B1-B7 亚型不同。2010 年 Maehongson 菌株的无毒成分基因表明,ha33、ha17 和 botR 基因与 Okra(B1)菌株接近,而 ha70 和 ntnh 基因与 111(B2)菌株接近。
结论/意义:本研究表明了对肉毒梭菌进行分子基因分型的有效性,以及它如何帮助我们了解肉毒毒素基因的流行病学和变异。因此,泰国最近发生的肉毒中毒事件是由不同的肉毒梭菌类型引起的。