通过CRISPR/Cas13辅助检测可检测耐卡巴尼姆的Klebsiella pneumoniae
Yaling Cao1, Yuan Tian1, Jing Huang2
1Beijing Institute of Hepatology/Beijing Youan Hospital, Capital Medical University, Beijing 100069, China.
概括
新的基于CRISPR的测定方法可以快速有效地检测抗卡巴因的Klebsiella肺炎 (CRKP). 这些方法改善了早期识别,使严重感染能够及时接受抗微生物治疗.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 传染性疾病 传染性疾病
背景情况:
- 耐卡巴胺的Klebsiella肺炎 (CRKP) 构成了严重的威胁,导致严重的社区和医院获得的感染.
- 目前用于CRKP识别的方法通常是复杂和耗时的,延迟了适当的治疗.
研究的目的:
- 开发用于早期检测CRKP的新高效方法.
- 为了使CRKP感染的及时和有效的抗微生物治疗.
主要方法:
- 设计的K. pneumoniae (KP),KPC和NDM特定的CRISPRRNAs (crRNAs),PCR原料和RAA原料.
- 已建立的CRISPR/Cas13a结合PCR (光) 和RAA (光和横流带) 试验.
- 使用61个临床菌株 (51个CRKP,10个对卡巴胺敏感) 的验证试验.
主要成果:
- 对于KP,blaKPC和blaNDM基因,PCR-CRISPR试验实现了1拷贝/μL的检测极限 (LOD).
- RAA-CRISPR试验实现了101副本/μL的LOD,具有光和侧流带读数.
- 对于目标基因,CRKP检测率为92.16% (47/51),具有高灵敏度和特异性 (100%).
结论:
- 开发了有效的PCR和RAA-CRISPR试验,用于检测碳酶基因 (blaKPC,blaNDM) 和KP.
- 这些测试有助于快速准确地识别CRKP.
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