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. 2023 Mar 1;85(3):329-333.
doi: 10.1292/jvms.22-0523. Epub 2023 Jan 18.

Simple and rapid detection of severe fever with thrombocytopenia syndrome virus in cats by reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay using a dried reagent

Keita Ishijima  1 , Kota Yokono  2 , Eunsil Park  1 , Masakatsu Taira  1 , Kango Tatemoto  1   3 , Yudai Kuroda  1 , Milagros Virhuez Mendoza  1   3 , Yusuke Inoue  1   3 , Michiko Harada  1   3 , Aya Matsuu  1 , Shigeru Morikawa  4 , Shuetsu Fukushi  5 , Ken Maeda  1   3
Affiliations

Simple and rapid detection of severe fever with thrombocytopenia syndrome virus in cats by reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay using a dried reagent

Keita Ishijima et al. J Vet Med Sci. .

Abstract

Severe fever with thrombocytopenia syndrome virus (SFTSV) causes lethal hemorrhagic diseases in human, cats, and dogs. Several human cases involving direct transmission of SFTSV from diseased animals have been reported. Therefore, rapid diagnosis in veterinary clinics is important for preventing animal-to-human transmission. Previously, we developed a simplified reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for human that does not require RNA extraction for detecting the SFTSV genome. In this study, we improved the simplified RT-LAMP assay for cats by introducing a dried reaction reagent and investigated the applicability of this method for diagnosing SFTS in cats. SFTSV RNA was detected in 11 of 12 cats naturally infected with SFTSV by RT-LAMP assay using both liquid and dried reagents. The RT-LAMP assay using liquid and dried reagents was also applicable to the detection of SFTSV genes 3-4 days after challenge in cats experimentally infected with SFTSV. The minimum copy number of SFTSV genes for 100% detection using the RT-LAMP assay with liquid and dried reagents was 4.3 × 104 and 9.6 × 104 copies/mL, respectively. Although the RT-LAMP assay using the dried reagent was less sensitive than that using the liquid reagent, it was sufficiently sensitive to detect SFTSV genes in cats with acute-phase SFTS. As the simplified RT-LAMP assay using a dried reagent enables detection of SFTSV genes more readily than the assay using a liquid reagent, it is applicable for use in veterinary clinics.

Keywords: cat; reverse transcription-loop-mediated isothermal amplification; reverse transcription-polymerase chain reaction; severe fever with thrombocytopenia syndrome virus; zoonosis.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig. 1.
Fig. 1.
Comparison of severe fever with thrombocytopenia syndrome virus (SFTSV) RNA copy number in serum and plasma of cats. Cats were divided into two groups based on the result (i.e., positive and negative) of reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay using liquid or dried reagent. The bars indicate the median value.
See this image and copyright information in PMC

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