{"title":"开发用于检测赤羽病毒的原位杂交方法","authors":"Nihat Toplu, T. Ç. Oğuzoğlu, A. Akkoç","doi":"10.14440/jbm.2024.413","DOIUrl":null,"url":null,"abstract":"Akabane virus (AKAV) is an arbovirus belonging to the family Bunyaviridae, genus Orthobunyavirus. AKAV consists of three-segment (L, M, and S RNA segments), negative single-stranded RNA. The aim of this study was to investigate an in situ hybridization method (ISH) in a Vero E6 cell line infected with Akabane virus. The 320 base pair amplicon was obtained by RT-PCR with a primer pair and labeled with digoxigenin. Akabane virus RNAs were seen as a granular pattern in the cytoplasm of infected cells. As a result, the expression of the particular Akabane virus gene area was successfully disclosed in the current investigation using the ISH method with a digoxigenin-labeled probe.","PeriodicalId":73618,"journal":{"name":"Journal of biological methods","volume":"17 3","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2024-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Development of an In Situ Hybridization Method for Detection of Akabane Virus\",\"authors\":\"Nihat Toplu, T. Ç. Oğuzoğlu, A. Akkoç\",\"doi\":\"10.14440/jbm.2024.413\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Akabane virus (AKAV) is an arbovirus belonging to the family Bunyaviridae, genus Orthobunyavirus. AKAV consists of three-segment (L, M, and S RNA segments), negative single-stranded RNA. The aim of this study was to investigate an in situ hybridization method (ISH) in a Vero E6 cell line infected with Akabane virus. The 320 base pair amplicon was obtained by RT-PCR with a primer pair and labeled with digoxigenin. Akabane virus RNAs were seen as a granular pattern in the cytoplasm of infected cells. As a result, the expression of the particular Akabane virus gene area was successfully disclosed in the current investigation using the ISH method with a digoxigenin-labeled probe.\",\"PeriodicalId\":73618,\"journal\":{\"name\":\"Journal of biological methods\",\"volume\":\"17 3\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-02-23\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of biological methods\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.14440/jbm.2024.413\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of biological methods","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.14440/jbm.2024.413","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
赤羽病毒（AKAV）是一种虫媒病毒，属于布尼亚病毒科、正布尼亚病毒属。AKAV 由三段（L、M 和 S RNA 段）阴性单链 RNA 组成。本研究的目的是在感染赤羽病毒的 Vero E6 细胞系中研究一种原位杂交方法（ISH）。使用引物对通过 RT-PCR 获得 320 碱基对扩增片段，并用地高辛标记。在感染细胞的细胞质中，可以看到赤羽病毒 RNA 呈颗粒状。因此，在本次研究中，使用地高辛标记探针的 ISH 方法成功地揭示了赤羽病毒特定基因区域的表达。
Development of an In Situ Hybridization Method for Detection of Akabane Virus
Akabane virus (AKAV) is an arbovirus belonging to the family Bunyaviridae, genus Orthobunyavirus. AKAV consists of three-segment (L, M, and S RNA segments), negative single-stranded RNA. The aim of this study was to investigate an in situ hybridization method (ISH) in a Vero E6 cell line infected with Akabane virus. The 320 base pair amplicon was obtained by RT-PCR with a primer pair and labeled with digoxigenin. Akabane virus RNAs were seen as a granular pattern in the cytoplasm of infected cells. As a result, the expression of the particular Akabane virus gene area was successfully disclosed in the current investigation using the ISH method with a digoxigenin-labeled probe.
京公网安备 11010802042870号
京ICP备2023020795号-1
分享
求助内容：
应助结果提醒方式：
扫码关注我们
