Accurate and sensitive analysis of Staphylococcus aureus through CRISPR-Cas12a based recycling signal amplification cascades for early diagnosis of skin and soft tissue infections

被引:21
|
作者
Wei, JingJing [1 ]
机构
[1] Zhuji Peoples Hosp Zhejiang Prov, Dept Dermatol, Zhuji City 311800, Zhejiang, Peoples R China
关键词
Skin and soft tissue infections; CRISPR-Cas12a; Staphylococcus aureus; PBP2a; BACTERIA; EPIDEMIOLOGY;
D O I
10.1016/j.mimet.2021.106167
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Early-diagnosis and treatments of skin and soft tissue infections remain a huge challenge due to the difficulties in the detection of trace amounts of bacteria. We develop here a novel method through CRISPR-Cas12a based recycling signal amplification cascades and demonstrated its feasibility of Staphylococcus aureus detection in a sensitive and accurate way. The highlights of the proposed method are calculated as: i) the designed allosteric probe is responsible for accurate identification of SA through PBP2a-specific aptamer; ii) high sensitivity from three processes, including DNA polymerase-based target SA release, Nb.BbvCI enzyme induced ssDNA generation and attached CRISPR-Cas12a. As a result, the proposed method exhibited a detection range from 10(6) to 10(2) CFU/ ml. Eventually, we believe that the proposed method could be expanded for the construction of diverse sensing platforms for detecting different trace bacteria.
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页数:5
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