The rapid and precise detection of pathogenic agents is critical for public health and societal stability. The detection of biological warfare agents (BWAs) is especially vital within military and counter-terrorism contexts, essential in defending against biological threats. Traditional methods, such as polymerase chain reaction (PCR), are limited by their need for specific settings, impacting their adaptability and versatility. This study introduces a cell-free biosensor for BWA detection by converting the 16S rRNA of targeted pathogens into detectable functional protein molecules. The modular nature of this approach allows for the flexible configuration of pathogen detection, enabling the simultaneous identification of multiple pathogenic 16S rRNAs through customized reporter proteins for each targeted sequence. Furthermore, we demonstrate how this method integrates with techniques utilizing retroreflective Janus particles (RJPs) for facile and highly sensitive pathogen detection. The cell-free biosensor, employing RJPs to measure the reflection of non-chromatic white light, can detect 16S rRNA from BWAs at femtomolar levels, corresponding to tens of colony-forming units per milliliter of pathogenic bacteria. These findings represent a significant advancement in pathogen detection, offering a more efficient and accessible alternative to conventional methodologies.

中文翻译：

---

一种用于生物战剂多重、灵敏检测的无细胞生物传感器

快速、准确地检测病原体对于公共卫生和社会稳定至关重要。生物战剂 (BWA) 的检测在军事和反恐背景下尤其重要，对于防御生物威胁至关重要。聚合酶链式反应 (PCR) 等传统方法因其对特定设置的需求而受到限制，从而影响了其适应性和多功能性。本研究引入了一种用于 BWA 检测的无细胞生物传感器，通过将目标病原体的 16S rRNA 转化为可检测的功能蛋白分子。这种方法的模块化性质允许病原体检测的灵活配置，从而能够通过每个目标序列的定制报告蛋白同时识别多种致病性 16S rRNA。此外，我们还演示了该方法如何与利用逆反射 Janus 粒子 (RJP) 的技术相结合，以实现轻松且高度灵敏的病原体检测。这种无细胞生物传感器采用 RJP 来测量非彩色白光的反射，可以以飞摩尔水平检测来自 BWA 的 16S rRNA，相当于每毫升病原菌数十个菌落形成单位。这些发现代表了病原体检测的重大进步，为传统方法提供了更有效、更容易使用的替代方案。