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

• Published in: Journal of Veterinary Medical Science Vol. 85; no. 3; pp. 329 - 333
• Main Authors: ISHIJIMA, Keita, YOKONO, Kota, PARK, Eunsil, TAIRA, Masakatsu, TATEMOTO, Kango, KURODA, Yudai, MENDOZA, Milagros Virhuez, INOUE, Yusuke, HARADA, Michiko, MATSUU, Aya, MORIKAWA, Shigeru, FUKUSHI, Shuetsu, MAEDA, Ken
• Format: Journal Article
• Language: English
• Published: Japan JAPANESE SOCIETY OF VETERINARY SCIENCE 2023; Japan Science and Technology Agency; The Japanese Society of Veterinary Science
• Subjects: Animals; cat; Cat Diseases; Cats; Copy number; Dog Diseases; Dogs; Fever; Gene amplification; Genomes; Hemorrhage; Humans; Indicators and Reagents; Nucleic Acid Amplification Techniques - methods; Nucleic Acid Amplification Techniques - veterinary; Phlebovirus - genetics; Reagents; Reverse transcription; reverse transcription-loop-mediated isothermal amplification; reverse transcription-polymerase chain reaction; RNA, Viral - genetics; Sensitivity and Specificity; Severe Fever with Thrombocytopenia Syndrome - veterinary; severe fever with thrombocytopenia syndrome virus; Thrombocytopenia; Veterinary services; Virology; zoonosis; reverse transcription-polymerase chain reaction; reverse transcription-loop-mediated isothermal amplification; zoonosis; cat; severe fever with thrombocytopenia syndrome virus
• DOI: 10.1292/jvms.22-0523
• ISSN: 0916-7250; 1347-7439

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.