マツイ　ユウト   Yuuto Matsui   松井　優人    所属   川崎医科大学  医学部 基礎医学　微生物学    職種   助教
論文種別	研究論文（学術雑誌）
言語種別	英語
査読の有無	査読あり
表題	Development of a Multi-Locus Real-Time PCR with a High-Resolution Melting Assay to Differentiate Wild-Type, Asian Recombinant, and Vaccine Strains of Lumpy Skin Disease Virus
掲載誌名	正式名：Veterinary Sciences 略　 称：Vet Sci.
掲載区分	国外
巻・号・頁	12(3)
国際共著	国際共著
著者・共著者	Kultyarat Bhakha, Yuto Matsui, Natchaya Buakhao, Saruda Wanganurakkul, Taweewat Deemagarn, Mami Oba, Hitoshi Takemae, Tetsuya Mizutani, Naoaki Misawa, Lerdchai Chintapitaksakul, Kentaro Yamada, Nutthakarn Suwankitwat
担当区分	筆頭著者
発行年月	2025/03
概要	Lumpy skin disease virus (LSDV) affects cattle and causes significant economic damage. The live vaccine derived from an attenuated strain is effective but is associated with mild disease and skin lesions in some vaccinated cattle. Moreover, recombinant LSDV strains, particularly one with wild-type field and vaccine strains, have recently emerged and spread throughout Asian countries. A cost-effective LSDV typing method is required. We developed a multi-locus real-time PCR with a high-resolution melting (HRM) assay to differentiate between the wild-type, vaccine, and dominant Asian recombinant strains. Based on a multiple alignment analysis, we selected three target genes for the HRM assay, ORF095, ORF126, and ORF145, in which there are insertions/deletions and nucleotide substitutions between wild-type and vaccine strains, and designed primer sets for the assay. Using the synthetic DNA encoding these genes for the two strains, it was shown that the PCR amplicons intercalated with a saturating fluorescent dye could clearly differentiate between wild-type and vaccine strains in the HRM analysis for all three target genes. Further, using clinical samples, our method was able to identify recombinant strains harboring the wild-type ORF095 and ORF145 and the vaccine strain ORF126 genes. Thus, our HRM assay may provide rapid LSDV typing.