Wang Xiaoyan, Hu Jinli
Department of Pediatrics, Affiliated Hospital of Shaoxing University, No 999 Zhongxing South Road, Yuecheng District, Shaoxing City, 312000, Zhejiang Province, China.
Biotechnol Lett. 2025 Jun 28;47(4):69. doi: 10.1007/s10529-025-03612-5.
Pseudomonas aeruginosa (P. aeruginosa) is a significant opportunistic pathogen associated with nosocomial infections, particularly in pediatric populations, where it can lead to severe clinical manifestations, including P. aeruginosa-associated meningitis. To meet the critical need for highly sensitive detection of P. aeruginosa, a novel fluorescent biosensor utilizing a three-way junction (TWJ) probe has been developed. This biosensor capitalizes on the specific binding interaction between P. aeruginosa and a tailored aptamer embedded within a DNA TWJ structure. Upon target binding, the double-stranded DNA branches of the TWJ undergo a conformational rearrangement, resulting in the formation of two distinct DNA "Y" junction structures. These structures are subsequently linked by a designed sequence, initiating a DNA polymerase/endonuclease-mediated strand displacement amplification process. The TWJ-based biosensor offers several key advantages: (i) the integration of the aptamer sequence within the TWJ probe ensures high specificity for target recognition, and (ii) the subsequent enzymatic amplification significantly enhances the sensitivity of detection. Under optimized experimental conditions, the biosensor demonstrated a broad linear detection range from 10 to 10 to 10 cfu/mL, with an exceptionally low limit of detection of 4.12 cfu/mL. Recovery studies further confirmed the reliability and robustness, highlighting its potential for clinical implementation. This innovative bio-sensing strategy represents a significant advancement in diagnostic technology, offering a promising tool for the early and accurate detection of infectious diseases in pediatric patients, with potential applications in improving clinical outcomes.
铜绿假单胞菌是一种重要的机会致病菌，与医院感染相关，尤其是在儿科人群中，它可导致严重的临床表现，包括铜绿假单胞菌相关性脑膜炎。为满足对铜绿假单胞菌进行高灵敏度检测的迫切需求，已开发出一种利用三链交界（TWJ）探针的新型荧光生物传感器。这种生物传感器利用了铜绿假单胞菌与嵌入DNA TWJ结构中的定制适配体之间的特异性结合相互作用。在靶标结合后，TWJ的双链DNA分支发生构象重排，导致形成两个不同的DNA“Y”型交界结构。这些结构随后通过设计的序列连接，启动DNA聚合酶/核酸内切酶介导的链置换扩增过程。基于TWJ的生物传感器具有几个关键优势：（i）TWJ探针内适配体序列的整合确保了对靶标识别的高特异性，（ii）随后的酶促扩增显著提高了检测灵敏度。在优化的实验条件下，该生物传感器的线性检测范围为10至10至10 cfu/mL，检测限低至4.12 cfu/mL。回收率研究进一步证实了其可靠性和稳健性，突出了其临床应用潜力。这种创新的生物传感策略代表了诊断技术的重大进步，为儿科患者传染病的早期准确检测提供了一种有前景的工具，在改善临床结果方面具有潜在应用。
