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. 2021 Apr 16;13(4):693.
doi: 10.3390/v13040693.

Development of an RT-LAMP Assay for the Rapid Detection of SFTS Virus

Shiori Sano  1 , Shuetsu Fukushi  2 , Souichi Yamada  2 , Shizuko Harada  2 , Hitomi Kinoshita  2 , Satoko Sugimoto  2 , Tomoki Yoshikawa  2 , Takeshi Kurosu  2 , Yuki Takamatsu  2 , Masayuki Shimojima  2 , Shoichi Toda  3 , Yuka Hamada  4 , Naoki Fujisawa  5 , Takayuki Sugimoto  6 , Masayuki Saijo  2
Affiliations

Development of an RT-LAMP Assay for the Rapid Detection of SFTS Virus

Shiori Sano et al. Viruses. .

Abstract

Detection of severe fever with thrombocytopenia syndrome (SFTS) virus (SFTSV) during the early phase of the disease is important for appropriate treatment, infection control, and prevention of further transmission. The reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a nucleic acid amplification method that amplifies the target sequence under isothermal conditions. Here, we developed an RT-LAMP with a novel primer/probe set targeting a conserved region of the SFTSV L segment after extraction of viral RNA (standard RT-LAMP). Both the Chinese and Japanese SFTSV strains, including various genotypes, were detected by the standard RT-LAMP. We also performed RT-LAMP using the same primer/probe set but without the viral RNA extraction step (called simplified RT-LAMP) and evaluated the diagnostic efficacy. The sensitivity and specificity of the simplified RT-LAMP were 84.9% (45/53) and 89.5% (2/19), respectively. The simplified RT-LAMP can detect SFTSV in human sera containing >103.5 copies/mL viral RNA. The two RT-LAMP positive but quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) negative samples were positive in the conventional RT-PCR, suggesting that there was no false positive reaction in the RT-LAMP. Both the standard and simplified RT-LAMP are useful for detecting the SFTSV genome in patients during the early phase of the disease.

Keywords: RT-LAMP; SFTS virus; simplified method.

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Conflict of interest statement

The authors declare no competing interest.

Figures

Figure 1
Figure 1
Alignment of SFTSV L segment sequences and positions of the primers used for RT-LAMP assay. The Genbank accession numbers for the L segment sequences of SFTSV strains are as follows, YG1: AB817979; SPL010: AB817983; SPL035: AB817986; SPL057: AB983500; SPL100:AB983519; SPL004:AB817981; SPL230; LC620253, JS4: HQ141604; SD4: HM802202, 2012YSH105: KF711869; WJ: HQ171186, and HB29: HM745930; SPL193; LC620254, SPL238; LC620252, SPL179; LC620255.
Figure 2
Figure 2
qPCR measurement of RNA copy numbers in serum samples that were positive and negative in the simplified RT-LAMP assay. There was a significant difference (p < 0.01) in RNA copy number in the positive and negative samples.
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References

    1. Yu X.-J., Liang M.-F., Zhang S.-Y., Liu Y., Li J.-D., Sun Y.-L., Zhang L., Zhang Q.-F., Popov V.L., Li C., et al. Fever with Thrombocytopenia Associated with a Novel Bunyavirus in China. N. Engl. J. Med. 2011;364:1523–1532. doi: 10.1056/NEJMoa1010095. - DOI - PMC - PubMed
    1. International Committee on Taxonomy of Viruses ICTV Virus Taxonomy: 2019 Release. [(accessed on 17 February 2021)]; Available online: https://talk.ictvonline.org/taxonomy/
    1. Li D. A highly pathogenic new bunyavirus emerged in China. Emerg. Microbes Infect. 2013;2:1–4. doi: 10.1038/emi.2013.1. - DOI - PMC - PubMed
    1. Hu J., Li Z., Cai J., Liu D., Zhang X., Jiang R., Guo X., Liu D., Zhang Y., Cui L., et al. A cluster of bunyavirus-associated severe fever with thrombocytopenia syndrome cases in a coastal plain area in China, 2015: Identification of a previously unidentified endemic region for severe fever with thrombocytopenia bunyavirus. Open Forum Infect. Dis. 2019;6:209. doi: 10.1093/ofid/ofz209. - DOI - PMC - PubMed
    1. Jung I.Y., Choi W., Kim J., Wang E., Park S.W., Lee W.J., Choi J.Y., Kim H.Y., Uh Y., Kim Y.K. Nosocomial person-to-person transmission of severe fever with thrombocytopenia syndrome. Clin. Microbiol. Infect. 2019;25:633.e1. doi: 10.1016/j.cmi.2019.01.006. - DOI - PubMed

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