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在具有挑战性的法医样本中对Yersinia pestis的类型化通过针对性下一代测序多部位变量数量并列重复区域

Hyeongseok Yun¹, Seung-Ho Lee¹, Se Hun Gu¹

¹5th Directorate, 3rd R&D Institute, Agency for Defense Development, Yeuseong P.O. Box 35, Daejeon 34186, Republic of Korea.

|October 29, 2025

概括

微生物法医现在可以在具有挑战性的样本中更好地识别像Yersinia pestis这样的细菌菌株. 一种新的有针对性的丰富方法提高了法医调查中多位置变量数并列重复分析 (MLVA) 的准确性.

科学领域:

微生物法医学微生物法医
分子生物学分子生物学
基因组学就是基因组学.

背景情况:

微生物法医分析武器化微生物的生物证据.
在模拟的法医样本中检测和类型化Yersinia pestis存在挑战.
全基因组测序 (WGS) 覆盖范围可能受到宿主DNA和低细菌丰度的限制.

研究的目的:

在模拟的法医样本中开发和评估一种准确的Yersinia pestis菌株类型的方法.
为了克服微生物法医学的WGS的局限性,特别是在痕迹或降解样本.

主要方法:

模拟的法医样本 (血,番茄汁,葡萄汁,手术口罩) 添加了Yersinia pestis.
实时PCR用于初始检测.
全基因组测序 (WGS) 和全基因组放大 (WGA),然后进行多位变量数串联重复分析 (MLVA).
为MLVA地点制定基于混合化的目标缩策略.

主要成果:

实时PCR证实了所有样本中的Yersinia pestis存在.
产品的WGS覆盖率差异很大 (0.46%97.1%),阻碍了菌株的识别.
基于WGA的MLVA分析显示精度低 (4%100%).
针对MLVA的目标丰富的NGS在所有样本中实现了100%的准确性,证实了单一菌株.

结论:

针对MLVA位点的向丰富是微生物法医学的有效方法.
这一策略克服了宿主DNA干扰和低微生物丰富度所带来的挑战.
该方法可以在复杂的法医样本中可靠地识别菌株水平.

关键词:

耶尔西尼亚瘟疫 (Yersinia pestis) 是一种传染病.法医微生物学法医微生物学多个位置的变量数量串联重复分析.有针对性的下一代测序.

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