Department of Dermatology, Zhuji Affiliated Hospital of Shaoxing University, No.9 Jianmin Road, Taozhu Street, Zhuji City, 311800, Zhejiang Province, China.
Mol Biotechnol. 2024 Jun;66(6):1402-1408. doi: 10.1007/s12033-023-00791-2. Epub 2023 Jun 26.
Staphylococcus aureus (S. aureus), as a Gram-positive bacterium, is commonly encountered in various infectious diseases, such as acute skin and soft tissue infections. Despite that many efforts have been made, sensitive and reliable quantitative determination of S. aureus remains a huge challenge. Here, we depict a novel colorimetric approach for sensitive and accurate detection by combining allosteric probe-based target recognition and chain extension-based dual signal recycling. The single-strand DNA (ssDNA) products generated by the chain extension process lead to the liberation of G-quadruplex sequences, which can fold into active DNAzyme under the assistance of hemin. The active DNAzyme can work as peroxidase mimics to catalyze the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS)-HO system, causing the color change of the system. Eventually, the method exhibits a wide detection range from 10 cfu/mL to 10 cfu/mL. The limit of detection of the approach was determined 232 cfu/mL. Considering the robust capability of the approach in S. aureus detection, we believe that it will be a potential alternative tool for biomedical research and clinical molecular diagnostics.
金黄色葡萄球菌(S. aureus)作为一种革兰氏阳性菌,常见于各种感染性疾病,如急性皮肤和软组织感染。尽管已经做了很多努力,但对金黄色葡萄球菌进行敏感和可靠的定量检测仍然是一个巨大的挑战。在这里,我们描述了一种新的比色法,通过结合变构探针靶标识别和基于链延伸的双重信号循环,实现了敏感和准确的检测。链延伸过程中产生的单链 DNA(ssDNA)产物导致 G-四链体序列的释放,在血红素的辅助下,G-四链体序列可以折叠成有活性的 DNA 酶。有活性的 DNA 酶可以作为过氧化物酶的模拟物,催化 2,2'-联氮双(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS)-HO 体系,引起体系颜色的变化。最终,该方法的检测范围从 10 cfu/mL 到 10 cfu/mL。该方法的检测限为 232 cfu/mL。鉴于该方法在金黄色葡萄球菌检测方面的强大能力,我们相信它将成为生物医学研究和临床分子诊断的一种有潜力的替代工具。