Wang Zhenfei, Zhang Jing, He Xiao, Wang Chunming, Li Zhenyue, Yang Zitong, Zhang Cheng, Fan Tingli, Su Kai
Institute of EcoHealth, School of Public Health, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China.
Department of Agricultural and Animal Husbandry Engineering, Cangzhou Technical College, Cangzhou, China.
Front Cell Infect Microbiol. 2025 Sep 2;15:1640503. doi: 10.3389/fcimb.2025.1640503. eCollection 2025.
The Respiratory Syncytial Virus (RSV) is a significant agent linked to respiratory infections, representing a considerable health risk for vulnerable populations, including infants, older adults, and those with weakened immune systems. This research successfully introduces an RNA extraction-free rapid detection technique for RSV utilizing real-time reverse transcription recombinase-aided amplification (RT-RAA) technology. Through the crafting of specific primers and probes, this approach enables precise identification of RSV without any interference from other prevalent respiratory viruses. Tests for sensitivity indicated that the detection threshold at a 95% confidence interval was 159 copies per reaction, while the visual detection limit was found to be 1,177 copies per reaction. Testing on clinical samples demonstrated a high degree of consistency with reverse transcription quantitative real-time PCR (RT-qPCR), achieving a Kappa value of 1, which signifies excellent correlation. Furthermore, the amplified products from RT-RAA can be seen with the aid of a portable blue light device, rendering this method appropriate for rapid detection in settings where resources are limited. A total of 265 clinical samples were tested, and the results showed 100% concordance with RT-qPCR. Compared with rapid antigen detection tests (RADTs), RT-RAA exhibited significantly higher sensitivity (100% . 93.8%). The rapid detection method for RSV using RT-RAA offers solid technical assistance for the early identification and prevention of RSV.
呼吸道合胞病毒（RSV）是一种与呼吸道感染相关的重要病原体，对包括婴儿、老年人和免疫系统较弱者在内的易感人群构成相当大的健康风险。本研究成功引入了一种利用实时逆转录重组酶辅助扩增（RT-RAA）技术的无RNA提取的RSV快速检测技术。通过设计特异性引物和探针，该方法能够精确鉴定RSV，不受其他常见呼吸道病毒的干扰。敏感性测试表明，在95%置信区间下的检测阈值为每个反应159个拷贝，而视觉检测限为每个反应1177个拷贝。对临床样本的检测显示与逆转录定量实时PCR（RT-qPCR）具有高度一致性，kappa值为1，表明相关性极佳。此外，RT-RAA的扩增产物可借助便携式蓝光设备观察到，这使得该方法适用于资源有限环境下的快速检测。共检测了265份临床样本，结果显示与RT-qPCR的一致性为100%。与快速抗原检测试验（RADTs）相比，RT-RAA表现出显著更高的敏感性（100%对93.8%）。使用RT-RAA的RSV快速检测方法为RSV的早期识别和预防提供了有力的技术支持。
