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在人皮样本中采用 Staphylococcus aureus DNA 的多部位序列排型方法

Hiroka Furuya¹, Kohei Ogura^2,3, Norihiko Takemoto⁴

¹Department of Clinical Laboratory Science, Faculty of Health Sciences, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan.

Microbiology and immunology

|August 14, 2023

概括

一种新的多重嵌套PCR方法可以准确地直接从皮肤DNA中类型化金黄色葡萄球菌 (S. aureus),即使存在其他细菌. 这种无培养方法成功地在治愈的压力损伤皮肤上识别了S. aureus序列类型.

科学领域:

微生物学微生物学
分子生物学分子生物学
皮肤病学皮肤病学

背景情况:

*金黄色葡萄球菌 (S. aureus) 常常殖民于皮肤和粘膜,特别是在医疗保健机构.
* 之前的研究表明,与正常皮肤相比,在治愈的压力损伤 (PI) 皮肤上,金黄色细菌的丰度更高.
* 传统的多部位序列类型 (MLST) 对微生物群DNA是不可靠的,原因是其他细菌物种的污染.

研究的目的:

* 开发一种新的多重嵌套PCR方法,直接从人类皮肤样本中获得准确的S. aureus MLST.
* 为了在临床样本中实现S. aureus的无培养类型化,克服污染问题.

主要方法:

* 设计了七对保存的黄金色菌特异性原料,用于多重嵌套PCR.
* 进行初始多重放大,然后在稀释样本上对MLST进行常规PCR.
* 对人类DNA和皮肤共生细菌的验证特异性.

主要成果:

* 该方法在有污染DNA的情况下,特别放大了S. aureus等位基因序列.
*从PI.治愈的皮肤的DNA中成功确定了S. aureus序列类型 (STs).
*已识别出属于克隆复合1 (CC1) 和CC5.5的具有主导性的STs.

结论:

*开发的多重嵌套PCR试验提供了一种可靠的无培养方法来定型S. aureus.
*这种方法可以应用于临床样本,可能简化S. aureus的检测和表征,而不需要细菌分离.

关键词:

黄金葡萄球菌黄金葡萄球菌临床样本临床样本没有文化的方法.多个位置的序列类型.多重复合PCR是一种复合PCR.压力损伤的压力损伤

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