Wang Yiwen, Huang Jing, Chen Meiyi, Jin Zha, Liu Tingting, Jiang Xi, Huang Rongsheng, Hu Kairao, Wei Zhengkai, Liu Quan
School of Animal Science and Technology, Foshan University, Foshan, 528225, Guangdong Province, China.
School of Animal Science and Technology, Foshan University, Foshan, 528225, Guangdong Province, China; College of Veterinary Medicine, Southwest University, Chongqing, 400715, China.
Microb Pathog. 2025 Feb;199:107251. doi: 10.1016/j.micpath.2024.107251. Epub 2024 Dec 20.
Beiji nairovirus (BJNV) is a recently discovered tick-borne RNA virus associated with human febrile illness. This study aimed to develop a loop-mediated isothermal amplification (LAMP) assay for the precise detection of BJNV, with a specific focus on assessing its effectiveness with clinical samples. The optimal molecular target was identified as the BJNV small (S) segment gene, and the ideal reaction conditions were established at 65 °C for 50 min. A neutral red stain concentration of 300 μM was determined to be optimal for visualizing the LAMP reaction. The LAMP method demonstrated an impressive lower limit of detection at 10 copies/μL, highlighting a sensitivity level 10,000 times higher than traditional PCR methods. Moreover, it showed no cross-reactivity with three other viruses. The LAMP assay demonstrated results consistent with those of semi-nested PCR when applied to clinical samples, thereby validating its suitability for field testing. In conclusion, the LAMP assay developed in this study represents a significant advancement in the rapid detection of BJNV. Its high sensitivity, specificity, and ease of use make it a valuable tool for the establishment of effective prevention and control strategies against this emerging pathogen.
贝吉内罗病毒（BJNV）是一种最近发现的与人类发热性疾病相关的蜱传RNA病毒。本研究旨在开发一种环介导等温扩增（LAMP）检测方法，用于精确检测BJNV，特别关注评估其在临床样本中的有效性。确定最佳分子靶标为BJNV小（S）片段基因，并确定在65°C下反应50分钟为理想反应条件。确定300μM的中性红染色浓度最适合用于观察LAMP反应。LAMP方法显示出令人印象深刻的10拷贝/μL的检测下限，突出了其比传统PCR方法高10000倍的灵敏度。此外，它与其他三种病毒没有交叉反应。当应用于临床样本时，LAMP检测结果与半巢式PCR结果一致，从而验证了其适用于现场检测。总之，本研究开发的LAMP检测方法代表了BJNV快速检测方面的重大进展。其高灵敏度、特异性和易用性使其成为制定针对这种新出现病原体的有效预防和控制策略的宝贵工具。
