Department of Infectious Diseases, Center of Infectious Diseases and Pathogen Biology, Key Laboratory of Organ Regeneration and Transplantation of the Ministry of Education, The First Hospital of Jilin University, Changchun, Jilin, China.
School of Basic Medicine, Inner Mongolia Medical University, Hohhot, Inner Mongolia, China.
Parasit Vectors. 2024 Nov 1;17(1):447. doi: 10.1186/s13071-024-06552-7.
Songling virus (SGLV) within the genus Orthonairovirus, family Nairoviridae, is an emerging tick-borne virus associated with human febrile illness. However, no rapid detection method for SGLV has been established.
In this study, four primer sets targeting the nucleocapsid protein gene of SGLV were designed for use in the LAMP assay and evaluated to identify the optimal primer set. Recombinant plasmids were constructed and utilized for assessing the sensitivity of the assay. Tacheng tick virus 1 (TcTV-1)-, Beiji nairovirus (BJNV)-, Yezo virus (YEZV)-, severe fever with thrombocytopenia syndrome virus (SFTSV)-, and tick-borne encephalitis virus (TBEV)-positive tick samples were utilized to assess the specificity. Field-collected ticks were also evaluated as biological specimens to validate the assay.
A SGLV-specific LAMP assay was established with a detection limit of 1 × 10 copies/μl and could be visually confirmed by a color change from purple to blue in SGLV-positive samples. No cross-reactivity was observed in the detection of TcTV-1, BJNV, YEZV, SFTSV, and TBEV using the LAMP assay. In addition to the detection of the same seven high-copy numbers of SGLV as the SYBR Green quantitative RT-PCR assay within a reduced timeframe, the developed LAMP method also effectively identified an additional sample with a low copy number in the field-collected tick samples.
We successfully developed a sensitive, specific, and cost-effective visual method for the rapid detection of SGLV using the LAMP assay, which can be applied in pathogenesis and epidemiological surveillance studies of SGLV.
Songling 病毒（SGLV）属于 Orthonairovirus 属、Nairoviridae 科，是一种与人类发热疾病相关的新兴蜱传病毒。然而，尚未建立 SGLV 的快速检测方法。
本研究设计了针对 SGLV 核衣壳蛋白基因的 4 对引物，用于 LAMP 检测，并对其进行了评估，以确定最佳引物对。构建了重组质粒，并用于评估检测方法的灵敏度。利用 Tacheng tick virus 1（TcTV-1）、Beiji nairovirus（BJNV）、Yezo virus（YEZV）、severe fever with thrombocytopenia syndrome virus（SFTSV）和 tick-borne encephalitis virus（TBEV）阳性蜱样本评估检测方法的特异性。还评估了野外采集的蜱作为生物样本，以验证该检测方法。
建立了一种 SGLV 特异性的 LAMP 检测方法，检测限为 1×10 拷贝/μl，在 SGLV 阳性样本中可通过颜色从紫色变为蓝色进行目视确认。LAMP 检测未观察到与 TcTV-1、BJNV、YEZV、SFTSV 和 TBEV 的交叉反应。除了在较短的时间内检测到与 SYBR Green 定量 RT-PCR 检测相同的七种高拷贝数的 SGLV 外，该开发的 LAMP 方法还能有效识别野外采集的蜱样本中的低拷贝数样本。
我们成功地开发了一种基于 LAMP 检测的敏感、特异、且经济有效的 SGLV 快速检测方法，可用于 SGLV 的发病机制和流行病学监测研究。
