Infectious diseases have become one of the most threatening global challenge with high morbidity and mortality, bringing great difficulties to clinical diagnosis and treatment. New strategy for high-specific and sensitive bacteria detection are urgently needed in facing the crisis of worldwide antibiotic resistance. Herein, a novel method through the integration of dual aptamer technology and CRISPR-Cas12a assisted rolling circle amplification (RCA) was present to obtain both accurate identification and high-sensitive detection of Methicillin-Resistant Staphylococcus Aureus (MRSA). The specificity inherited from the dual functionalized aptamers initiated bioconjugation to specifically recognize the protein targets on the surface of bacteria. Besides the target activity, the functionalized aptamer could also convert the protein recognition to nucleic acids signals. Through the integration of attached RCA and CRISPR-Cas12a assisted trans-cleavage, dual amplification of the nucleic acid signal was obtained. Based on this, we have extended the application of CRISPR-Cas12a from the nucleic acid detection to bacteria detection. As a result, the proposed method was demonstrated to be with significantly improved sensitivity towards MRSA detection. We believe that the novel integrated strategy would diversify the existing pool of bacterial detection and inspire the development of promising drug candidates in the future.

中文翻译：

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通过双功能适体和CRISPR-Cas12a辅助滚环扩增进行准确的MRSA鉴定。

传染病以高发病率和高死亡率成为全球最具挑战性的挑战之一，给临床诊断和治疗带来了巨大困难。面对全球抗生素耐药性危机，迫切需要一种用于检测高特异性和敏感细菌的新策略。在本文中，提出了一种通过双重适体技术和CRISPR-Cas12a辅助滚环扩增（RCA）的集成来获得耐甲氧西林金黄色葡萄球菌（MRSA）的准确鉴定和高灵敏度检测的新方法。从双功能适体继承的特异性启动了生物缀合，以特异性识别细菌表面上的蛋白质靶标。除了目标活动之外 功能化的适体还可以将蛋白质识别转换为核酸信号。通过结合的RCA和CRISPR-Cas12a辅助的反式切割的整合，获得了核酸信号的双重扩增。基于此，我们将CRISPR-Cas12a的应用范围从核酸检测扩展到了细菌检测。结果，证明了所提出的方法对MRSA检测的灵敏度大大提高。我们相信，这种新颖的综合策略将使现有的细菌检测池多样化，并激发未来有希望的候选药物的开发。我们已经将CRISPR-Cas12a的应用从核酸检测扩展到细菌检测。结果，证明了所提出的方法对MRSA检测的灵敏度大大提高。我们相信，这种新颖的综合策略将使现有的细菌检测池多样化，并激发未来有希望的候选药物的开发。我们已经将CRISPR-Cas12a的应用从核酸检测扩展到细菌检测。结果，证明了所提出的方法对MRSA检测的灵敏度大大提高。我们相信，新的整合策略将使现有的细菌检测池多样化，并激发未来有希望的候选药物的开发。