Department of Neonatology, The First Hospital of China Medical University, Shenyang City, Liaoning Province, 110000, China.
Appl Biochem Biotechnol. 2024 Oct;196(10):6716-6725. doi: 10.1007/s12010-024-04862-1. Epub 2024 Feb 24.
In the field of neonatal infections nursing, methicillin-resistant Staphylococcus aureus (MRSA) is a major bacterial pathogen. Here, we present a portable biosensor for MRSA detection that is both highly sensitive and portable, owing to its implementation on the personal glucose meter (PGM) platform. The H probe was fixed on the magnetic bead for mecA gene analysis. A blunt 3' terminus appeared in the MBs-H probe when the mecA gene was present. Exonuclease-III (Exo-III) recognized the blunt terminus and cleaved it, freeing the mecA gene and so facilitating target recycling. In the meantime, the remaining H probe-initiated hybridization chain reaction (HCR) led to the desired signal amplification. Portable quantitative detection of mecA gene is possible because PGM can read the quantity of invertase tagged on HCR product. After optimizing several experimental parameters, such as the concentration of Exo-III and incubation time, the constructed sensor is extremely sensitive, with a detection limit of 2 CFU/mL. The results from this sensitive PGM-based sensor are in agreement with those obtained from plate counting methods, suggesting that it can be used to accurately assess the MRSA content in artificial clinical samples. In addition, the PGM sensor can significantly cut down on time spent compared to plate counting techniques. The manufactured sensor provides a promising option for accurate identification of pathogenic bacteria.
在新生儿感染护理领域，耐甲氧西林金黄色葡萄球菌（MRSA）是一种主要的细菌病原体。在这里，我们提出了一种基于个人血糖仪（PGM）平台的高灵敏度和便携性的 MRSA 检测便携式生物传感器。H 探针被固定在磁珠上，用于 mecA 基因分析。当 mecA 基因存在时，MBs-H 探针上出现平头 3'末端。外切酶-III（Exo-III）识别平头末端并将其切割，释放 mecA 基因，从而促进靶标循环。同时，剩余的 H 探针启动杂交链式反应（HCR），导致所需的信号放大。由于 PGM 可以读取 HCR 产物上标记的 invertase 的数量，因此可以实现对 mecA 基因的便携式定量检测。在优化了 Exo-III 的浓度和孵育时间等几个实验参数后，构建的传感器具有极高的灵敏度，检测限低至 2 CFU/mL。该敏感的基于 PGM 的传感器的结果与平板计数方法获得的结果一致，表明它可用于准确评估人工临床样本中的 MRSA 含量。此外，PGM 传感器与平板计数技术相比，可显著缩短检测时间。制造的传感器为准确鉴定病原菌提供了一种很有前途的选择。
