4.3 Article

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

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JOURNAL OF MICROBIOLOGICAL METHODS
卷 183, 期 -, 页码 -

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ELSEVIER
DOI: 10.1016/j.mimet.2021.106167

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Skin and soft tissue infections; CRISPR-Cas12a; Staphylococcus aureus; PBP2a

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In this study, a novel method using CRISPR-Cas12a for sensitive and accurate detection of Staphylococcus aureus was developed, with high sensitivity and accuracy demonstrated. The proposed method has a potential for diverse sensing platforms for detecting different trace bacteria.
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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