2025/05/08 by Wenlong Huang, Meiyi Chen, Yiwen Wang +7 · 1 voice · 5 citations
Agricultural and Biological Sciences · Chemistry · Medicine · #Biology #Chemistry #Chromatography #DNA #Detection limit #GenBank #Gene #Genetics #Loop-mediated isothermal amplification #Molecular biology #Mosquito-borne diseases and control #Plant Virus Research Studies #Polymerase chain reaction #Real-time polymerase chain reaction #Reverse Transcription Loop-mediated Isothermal Amplification #Reverse transcriptase #SYBR Green I #Viral Infections and Vectors #Virology #Virus
paper · doi:10.1099/jgv.0.002094
published in Journal of General Virology 106(5) (Microbiology Society)
openalex publication_date 2025/05/08 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/26
Alongshan virus (ALSV) is a recently discovered tick-borne zoonotic virus. Currently, there is no rapid and accurate clinical method for ALSV detection. This study aimed to develop a loop-mediated isothermal amplification (LAMP) assay for precise ALSV infection detection. Specific primers were designed based on the S1 segment of the ALSV NE-TH4 strain’s genome (GenBank accession no. ON408067.1). The reaction time, temperature and concentration of the neutral red staining solution in the LAMP assay were optimized. Thorough evaluations of specificity, sensitivity and repeatability led to the development of a visually interpretable LAMP assay. The optimal amplification time was 50 min. The minimum detection limit for cDNA was as low as 0.005 pg μl −1 , and sensitivity for standards was 1.68×10 3 copies per μl, surpassing that of PCR and real-time PCR. No cross-reactivity was observed with Jingmen tick virus, Bole tick virus 4 and Beiji nairovirus. These results indicate that the LAMP assay is more sensitive and accurate than PCR and real-time PCR. The developed LAMP assay allows for on-site detection, reduces testing costs and provides rapid and accurate results. Thus, it lays a solid foundation for the prevention and control of emerging tick-borne ALSV.