Development of a LAMP assay for the rapid visual detection of the emerging tick-borne Songling virus.

IF 3 2区医学 Q1 PARASITOLOGY Parasites & Vectors Pub Date : 2024-11-01 DOI:10.1186/s13071-024-06552-7

Zheng Gui, Yuanning Ren, Qiqi Guo, Weiying Yang, Ziyan Liu, Ning Liu, Yunzhi Peng, Yu Liu, Jingfeng Yu, Lichao Sun, Zedong Wang

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Abstract

Background: 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.

Methods: 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.

Results: A SGLV-specific LAMP assay was established with a detection limit of 1 × 10^-2 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.

Conclusions: 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.

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开发用于快速肉眼检测新出现的蜱传松岭病毒的 LAMP 检测方法。

背景：松岭病毒（SGLV）是一种新出现的蜱传病毒，与人类发热性疾病有关。然而，目前还没有建立 SGLV 的快速检测方法：本研究设计了四组针对 SGLV 核壳蛋白基因的引物用于 LAMP 检测，并对其进行了评估，以确定最佳引物组。构建了重组质粒，并用于评估该检测方法的灵敏度。利用塔城蜱病毒 1（TcTV-1）、北吉奈洛病毒（BJNV）、野蜱病毒（YEZV）、严重发热伴血小板减少综合征病毒（SFTSV）和蜱传脑炎病毒（TBEV）阳性蜱样本来评估特异性。此外，还将野外采集的蜱虫作为生物标本进行评估，以验证该检测方法：结果：建立了一种 SGLV 特异性 LAMP 检测方法，其检测限为 1 × 10-2 拷贝/μl。使用 LAMP 法检测 TcTV-1、BJNV、YEZV、SFTSV 和 TBEV 时未发现交叉反应。除了能在更短的时间内检测出与 SYBR Green 定量 RT-PCR 检测方法相同的 7 种高拷贝数 SGLV 外，所开发的 LAMP 方法还能在野外采集的蜱虫样本中有效地鉴定出另外一种低拷贝数样本：我们成功地开发出了一种灵敏、特异且经济高效的可视化方法，利用 LAMP 检测法快速检测 SGLV，该方法可用于 SGLV 的致病机理和流行病学监测研究。

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来源期刊

Parasites & Vectors 医学-寄生虫学

CiteScore

6.30

自引率

9.40%

发文量

433

审稿时长

1.4 months

期刊介绍： Parasites & Vectors is an open access, peer-reviewed online journal dealing with the biology of parasites, parasitic diseases, intermediate hosts, vectors and vector-borne pathogens. Manuscripts published in this journal will be available to all worldwide, with no barriers to access, immediately following acceptance. However, authors retain the copyright of their material and may use it, or distribute it, as they wish. Manuscripts on all aspects of the basic and applied biology of parasites, intermediate hosts, vectors and vector-borne pathogens will be considered. In addition to the traditional and well-established areas of science in these fields, we also aim to provide a vehicle for publication of the rapidly developing resources and technology in parasite, intermediate host and vector genomics and their impacts on biological research. We are able to publish large datasets and extensive results, frequently associated with genomic and post-genomic technologies, which are not readily accommodated in traditional journals. Manuscripts addressing broader issues, for example economics, social sciences and global climate change in relation to parasites, vectors and disease control, are also welcomed.