A novel multilocus variable-number tandem repeat analysis for Bordetella parapertussis
A novel multilocus variable-number tandem repeat analysis for Bordetella parapertussis
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副百日咳博德特氏菌的新型多位点可变数量串联重复分析
DOI：
10.1099/jmm.0.001095
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发表时间：
2019
影响因子：
3
通讯作者：
Watanabe Mineo
中科院分区：
文献类型：
--
作者：
Kamachi Kazunari;Otsuka Nao;Fumimoto Rei;Ozawa Kensuke;Yao Shu-Man;Chiang Chuen-Sheue;Luu Laurence Don Wai;Lan Ruiting;Shibayama Keigo;Watanabe Mineo
目的：人类适应性副百日咳博德特氏菌是百日咳的病原体之一;然而，目前还没有对这种细菌病原体具有高分辨力的基因分型系统。因此，我们的目标是开发一个多位点可变数目串联重复序列分析（MLVA）的人类adapteddB。方法学：在副百日咳杆菌中有四个高度多态性的可变数目串联重复序列（VNTR）基因座。选择副百日咳菌基因组，采用多重PCR扩增。基于VNTR基因座处的串联重复的数目进行MLVA。用3株实验室参考菌株和50株人源B.结果：多重PCR为基础的MLVA表征53 B。将副百日咳参考菌株和分离株分为25个MLVA型，Simpson多样性指数为0.91（95%可信区间0.86-0.97）。三种参考菌株表现出不同的MLVA类型。31株日本分离株、10株法国分离株和3株台湾分离株分别属于14、9和3个MLVA型。相比之下，所有五个澳大利亚分离株属于同一类型。从已知流行病学联系的患者中收集的两种日本分离株具有相同的类型。结论。我们的新型MLVA方法对人类B基因分型具有很高的区分能力。副百日咳。关于这种生物体，这种基因分型系统是流行病学监测和调查疫情的一个有前途的工具。
Purpose.Human-adaptedBordetella parapertussisis one of the causative agents of whooping cough; however, there are currently no genotyping systems with high discriminatory power for this bacterial pathogen. We therefore aimed to develop a multilocus variable-number tandem repeat analysis (MLVA) for human-adaptedB. parapertussis.Methodology.Four highly polymorphic variable number tandem repeat (VNTR) loci in theB. parapertussisgenome were selected and amplified by multiplex PCR. MLVA was performed based on the number of tandem repeats at VNTR loci. The discriminatory power of MLVA was evaluated with three laboratory reference strains and 50 human isolates ofB. parapertussis.Results.Multiplex PCR-based MLVA characterized 53B. parapertussisreference strains and isolates into 25 MLVA types and the Simpson diversity index was 0.91 (95 % confidence interval, 0.86–0.97). The three reference strains exhibited different MLVA types. Thirty-one Japanese isolates, ten French isolates and three Taiwanese isolates belonged to fourteen, nine and three MLVA types, respectively. In contrast, all five Australian isolates belonged to the same type. Two Japanese isolates collected from patients with known epidemiological links had the same type.Conclusion.Our novel MLVA method has high discriminatory power for genotyping humanB. parapertussis. Regarding this organism, this genotyping system is a promising tool for epidemiological surveillance and investigating outbreaks.