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Unraveling potential gene biomarkers for dengue infection through RNA sequencing. 通过 RNA 测序揭示登革热感染的潜在基因生物标志物。
IF 1.9 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-10-14 DOI: 10.1007/s11262-024-02114-2
Jeyanthi Suppiah, Saiful Safuan Md Sani, Safiah Sabrina Hassan, Nur Iman Fasohah Nadzar, Nurul 'Izzah Ibrahim, Ravindran Thayan, Rozainanee Mohd Zain

Dengue virus hijacks host cell mechanisms and immune responses in order to replicate efficiently. The interaction between the host and the virus affects the host's gene expression, which remains largely unexplored. This pilot study aimed to profile the host transcriptome as a potential strategy for identifying specific biomarkers for dengue prediction and detection. High-throughput RNA sequencing (RNA-seq) was employed to generate host transcriptome profiles in 16 dengue patients and 10 healthy controls. Differentially expressed genes (DEGs) were identified in patients with severe dengue and those with dengue with warning signs compared to healthy individuals. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to elucidate the functions of upregulated and downregulated genes. Compared to healthy controls, 6466 genes were significantly differentially expressed (p < 0.05) in the dengue with warning signs group and 3082 genes in the severe dengue group, with over half being upregulated. The major KEGG pathways implicated included transport and catabolism (14.4%-16.3%), signal transduction (6.6%-7.3%), global and overview maps (6.7%-7.1%), viral diseases (4.6%-4.8%), and the immune system (4.4%-4.6%). Several genes exhibited consistent and significant upregulation across all dengue patients, regardless of severity: Interferon alpha inducible protein 27 (IFI27), Potassium Channel Tetramerization Domain Containing 14 (KCTD14), Syndecan 1 (SDC1), DCC netrin 1 receptor (DCC), Ubiquitin C-terminal hydrolase L1 (UCHL1), Marginal zone B and B1 cell-specific protein (MZB1), Nestin (NES), C-C motif chemokine ligand 2 (CCL2), TNF receptor superfamily member 17 (TNFSF17), and TNF receptor superfamily member 13B (TNFRSF13B). Further analysis revealed potential biomarkers for severe dengue prediction, including TNF superfamily member 15 (TNFSF15), Plasminogen Activator Inhibitor-2 (SERPINB2), motif chemokine ligand 7 (CCL7), aconitate decarboxylase 1 (ACOD1), Metallothionein 1G (MT1G), and Myosin Light Chain Kinase (MYLK2), which were expressed 3.5 times, 2.9 times, 2.3 times, 2.1 times, 1.7 times, and 1.4 times greater, respectively, than dengue patients exhibiting warning signs. The identification of these host biomarkers through RNA-sequencing holds promising implications and potential to augment existing dengue detection algorithms, contributing significantly to improved diagnostic and prognostic capabilities.

登革热病毒劫持宿主细胞机制和免疫反应,以便高效复制。宿主与病毒之间的相互作用会影响宿主的基因表达,而这一点在很大程度上仍未得到探索。这项试验性研究旨在分析宿主转录组,作为确定登革热预测和检测特异性生物标志物的潜在策略。研究人员采用高通量 RNA 测序(RNA-seq)技术生成了 16 名登革热患者和 10 名健康对照者的宿主转录组图谱。与健康人相比,在严重登革热患者和有登革热预警征兆的登革热患者中发现了差异表达基因(DEGs)。通过基因本体(GO)和京都基因和基因组百科全书(KEGG)分析,阐明了上调和下调基因的功能。与健康对照组相比,6466 个基因有明显的表达差异(p
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引用次数: 0
Domestic cat hepadnavirus genotype B is present in Southern Brazil. 巴西南部存在家猫狂犬病毒基因 B 型。
IF 1.9 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-10-14 DOI: 10.1007/s11262-024-02115-1
Alaíse Tessmann, Juliana Sumienski, Alexandre Sita, Larissa Mallmann, Gabriela Espíndola Birlem, Nilson Júnior da Silva Nunes, Camila Gottlieb Lupion, Juliana Schaeffer Eckert, Meriane Demoliner, Juliana Schons Gularte, Paula Rodrigues de Almeida, Fernando Rosado Spilki, Matheus Nunes Weber

Domestic cat hepadnavirus (DCH) (Orthohepadnavirus felisdomestici) is an emerging virus related to the hepatitis B virus (HBV) already reported in many countries. The molecular prevalence of DCH varies widely in the regions investigated so far. In the present work, we reported the presence of DCH in Brazil. Sixty cat serum samples tested by DCH presence using PCR and 1.67% (1/60) were positive, similar to the low positive molecular rates reported in United States and Japan. The DCH full-length genome was classified in genotype B, which is uncommon since this genotype was only reported once in Japan. The DCH-positive sample was obtained in a stray cat female apparently healthy, presenting ALT, AST, and ALKP normal values, and negative for FIV and FeLV. Due the low positivity rate detected, some factors as alteration in hepatic enzymes and FIV/FeLV infection could not be evaluated. Other works are necessary to statistically validate these observations in Brazil.

家猫细小病毒(DCH)(Orthohepadnavirus felisdomestici)是一种新出现的病毒,与乙型肝炎病毒(HBV)有关,已在许多国家报道。迄今为止,在所调查的地区中,DCH 的分子流行率差异很大。在本研究中,我们报告了巴西存在 DCH 的情况。我们使用 PCR 对 60 份猫血清样本进行了 DCH 检测,结果显示 1.67%(1/60)呈阳性,与美国和日本报告的较低分子阳性率相似。DCH 全长基因组被归类为基因型 B,这并不常见,因为这种基因型只在日本报道过一次。DCH 阳性样本来自一只流浪猫,该猫雌性,表面健康,ALT、AST 和 ALKP 值正常,FIV 和 FeLV 阴性。由于检测到的阳性率较低,因此无法对肝酶变化和 FIV/FeLV 感染等因素进行评估。有必要在巴西开展其他工作,对这些观察结果进行统计验证。
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引用次数: 0
The isolation and serotyping of foot-and-mouth disease virus in Iran during 2019-2022. 2019-2022 年伊朗口蹄疫病毒的分离和血清分型。
IF 1.9 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-10-12 DOI: 10.1007/s11262-024-02116-0
Siamak Khoshnood, Seyed Mahmoud Azimi, Zahra Ziafati Kafi, Hamideh Najafi, Arash Ghalyanchilangeroudi

Foot-and-mouth disease (FMD) is a significant transboundary animal disease that has a considerable economic impact on livestock systems worldwide. In order to determine the presence and type of FMD virus in Iran, a total of 90 samples of vesicular fluid and epithelial tissue were collected from the tongues, tooth pads, and hooves of clinically suspect cattle on 40 vaccinated farms in 9 provinces of Iran. These samples were collected during four years, from January 2019 to December 2022, and the vaccine was a locally produced polyvalent inactivated vaccine. The collected samples were analyzed using ELISA and isolation methods to identify and characterize the FMD virus. The results of the ELISA tests revealed that 66.66% of the samples were positive for FMD, and the serotypes of the virus were determined. Considering ELISA reslut, 62% of the samples were assigned to serotype O, 33% to serotype A, and 5% to serotype Asia-1. Furthermore, 90% of the positive samples were inoculated onto monolayer cultures of pig kidneys (IB-RS2) for isolation and antigen detection by serotype-specific ELISA kit. The great majority of detected serotype O viruses were from Esfahan province, while the most detected serotype A and serotype Asia-1 viruses were from Qom and Tehran provinces, respectively. These findings indicate that the ELISA and isolation methods are suitable for identifying and typing FMD viruses. The vaccination program in Iran, which includes three serotypes (O, A, and Asia-1), appears to be effective in controlling the spread of the disease. However, the continued circulation of these serotypes in most provinces suggests that ongoing surveillance and vaccination efforts are necessary.

口蹄疫(FMD)是一种严重的跨境动物疫病,对全球畜牧业系统造成了巨大的经济影响。为了确定伊朗是否存在口蹄疫病毒及其类型,我们从伊朗 9 个省 40 个疫苗接种农场的临床可疑牛的舌头、齿垫和蹄中收集了共计 90 份水泡液和上皮组织样本。这些样本是在 2019 年 1 月至 2022 年 12 月的四年期间收集的,疫苗是当地生产的多价灭活疫苗。采集的样本采用酶联免疫吸附试验(ELISA)和分离法进行分析,以确定口蹄疫病毒的身份和特征。ELISA 检测结果显示,66.66% 的样本对口蹄疫病毒呈阳性反应,病毒血清型也已确定。根据酶联免疫吸附试验的结果,62%的样本属于血清型 O,33%属于血清型 A,5%属于血清型 Asia-1。此外,90%的阳性样本被接种到猪肾单层培养物(IB-RS2)上,通过血清型特异性 ELISA 试剂盒进行分离和抗原检测。检测到的绝大多数血清型 O 病毒来自伊斯法罕省,而检测到最多的血清型 A 和血清型亚洲-1 病毒分别来自库姆省和德黑兰省。这些结果表明,酶联免疫吸附法和分离法适用于对口蹄疫病毒进行鉴定和分型。伊朗的疫苗接种计划包括三种血清型(O 型、A 型和亚洲-1 型),似乎能有效控制疾病的传播。然而,这些血清型在大多数省份的持续流行表明,有必要持续开展监测和疫苗接种工作。
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引用次数: 0
Identification and epidemiological analysis of a putative novel hantavirus in Australian flying foxes. 澳大利亚飞狐中一种假定的新型汉坦病毒的鉴定和流行病学分析。
IF 1.9 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-10-11 DOI: 10.1007/s11262-024-02113-3
Craig S Smith, Darren J Underwood, Anita Gordon, Michael J Pyne, Anna Smyth, Benjamin Genge, Luke Driver, David G Mayer, Jane Oakey

In July 2017, an investigation into the cause of neurological signs in a black flying fox (Pteropus alecto, family Pteropodidae) identified a putative novel hantavirus (Robina virus, ROBV, order Bunyavirales, family Hantaviridae, genus Mobatvirus) in its brain. Analysis of the evolutionary relationship between other hantaviruses using maximum-likelihood, a systematic Bayesian clustering approach, and a minimum spanning tree, all suggest that ROBV is most closely related to another Mobatvirus, Quezon virus, previously identified in the lung of a Philippine frugivorous bat (Rousettus amplexicaudatus, also family Pteropodidae). Subsequently, between March 2018 and October 2023, a total of 495 bats were opportunistically screened for ROBV with an experimental qRT-PCR. The total prevalence of ROBV RNA detected in Pteropus spp. was 4.2% (95% CI 2.8-6.4%). Binomial modelling identified that there was substantial evidence supporting an increase (P = 0.033) in the detection of ROBV RNA in bats in 2019 and 2020 suggesting of a possible transient epidemic. There was also moderate evidence to support the effect of season (P = 0.064), with peak detection in the cooler seasons, autumn, and winter, possibly driven by physiological and ecological factors similar to those already identified for other bat-borne viruses. This is Australia's first reported putative hantavirus and its identification could expand the southern known range of hantaviruses in Australasia.

2017年7月,一项针对黑飞狐(Pteropus alecto,翼足目翼足科)神经症状原因的调查在其大脑中发现了一种推测的新型汉坦病毒(Robina virus,ROBV,Bunyavirales目,Hantaviridae科,Mobatvirus属)。使用最大似然法、系统贝叶斯聚类方法和最小生成树分析其他汉坦病毒之间的进化关系,均表明 ROBV 与另一种 Mobatvirus(奎松病毒)关系最为密切,奎松病毒之前在菲律宾食俭蝙蝠(Rousettus amplexicaudatus,也是翼手目蝙蝠科)的肺部中被发现。随后,在 2018 年 3 月至 2023 年 10 月期间,利用实验性 qRT-PCR 对 495 只蝙蝠进行了 ROBV 的机会性筛查。在翼手目蝙蝠中检测到的 ROBV RNA 总流行率为 4.2%(95% CI 2.8-6.4%)。二项式建模发现,有大量证据支持 2019 年和 2020 年蝙蝠中 ROBV RNA 的检测率上升(P = 0.033),这表明可能会出现短暂流行。也有中等程度的证据支持季节的影响(P = 0.064),在凉爽的季节、秋季和冬季出现检测高峰,这可能是受生理和生态因素的驱动,类似于其他蝙蝠传播病毒已确定的因素。这是澳大利亚首次报告的假定汉坦病毒,对它的鉴定可能会扩大汉坦病毒在澳大拉西亚南部的已知范围。
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引用次数: 0
Molecular epidemiology and genetic evolution of avian influenza H5N1 subtype in Nigeria, 2006 to 2021. 2006 年至 2021 年尼日利亚 H5N1 亚型禽流感的分子流行病学和基因演变。
IF 1.9 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-10-01 Epub Date: 2024-06-19 DOI: 10.1007/s11262-024-02080-9
Ridwan O Adesola, Bernard A Onoja, Andrew M Adamu, Sheriff T Agbaje, Modinat D Abdulazeez, Olalekan C Akinsulie, Adetolase Bakre, Oyelola A Adegboye

Nigeria recorded one of the earliest outbreaks of the Highly Pathogenic Avian Influenza (HPAI) virus H5N1 in 2006, which spread to other African countries. In 2023, 18 countries reported outbreaks of H5N1 in poultry, with human cases documented in Egypt, Nigeria, and Djibouti. There is limited information on the molecular epidemiology of HPAI H5N1 in Nigeria. We determined the molecular epidemiology and genetic evolution of the virus from 2006 to 2021. We investigated the trend and geographical distribution across Nigeria. The evolutionary history of 61 full-length genomes was performed from 13 countries worldwide, and compared with sequences obtained from the early outbreaks in Nigeria up to 2021. MEGA 11 was used to determine the phylogenetic relationships of H5N1 strains, which revealed close ancestry between sequences in Nigeria and those from other African countries. Clade classification was performed using the subspecies classification tool for Bacterial and Viral Bioinformatics Research Center (BV-BRC) version 3.35.5. H5N1 Clade 2.2 was observed in 2006, with 2.3.2, 2.3.2.1f clades observed afterwards and 2.3.4.4b in 2021. Our findings underscore the need for genomics surveillance to track antigenic variation and clades switching to monitor the epidemiological of the virus and safeguard human and animal health.Impacts Specific variations in the hemagglutinin (HA) and neuraminidase (NA) genes of Avian influenza virus are consistent in different geographical regions. H5N1 Clade 2.2 was reported in 2006, with 2.3.2, 2.3.2.1f afterwards and 2.3.4.4b in 2021. Nigeria is an epicentre for avian influenza with three major migratory routes for wild birds transversing the country. It is plausible that the Avian influenza in Northern Nigeria may be linked to wild bird sanctuaries in the region.

尼日利亚是 2006 年最早爆发高致病性禽流感病毒 H5N1 的国家之一,该病毒随后蔓延到其他非洲国家。2023 年,18 个国家报告了家禽中 H5N1 病毒的爆发,埃及、尼日利亚和吉布提记录了人类病例。有关尼日利亚高致病性禽流感 H5N1 分子流行病学的信息十分有限。我们确定了 2006 年至 2021 年期间病毒的分子流行病学和基因演变情况。我们调查了尼日利亚的趋势和地理分布。我们对来自全球 13 个国家的 61 个全长基因组的进化史进行了研究,并与从尼日利亚早期疫情爆发到 2021 年期间获得的序列进行了比较。利用 MEGA 11 确定了 H5N1 菌株的系统发育关系,结果显示尼日利亚的序列与其他非洲国家的序列具有近亲关系。使用细菌和病毒生物信息学研究中心(BV-BRC)3.35.5 版的亚种分类工具进行了支系分类。2006 年观察到 H5N1 2.2 支系,之后观察到 2.3.2 和 2.3.2.1f 支系,2021 年观察到 2.3.4.4b 支系。我们的研究结果表明,有必要进行基因组学监测,以跟踪抗原变异和支系切换情况,从而监测病毒的流行病学,保障人类和动物的健康。2006 年报告了 H5N1 2.2 支系,之后报告了 2.3.2、2.3.2.1f 和 2.3.4.4b 支系。尼日利亚是禽流感的中心,有三条主要的野生鸟类迁徙路线横穿该国。尼日利亚北部的禽流感可能与该地区的野生鸟类保护区有关。
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引用次数: 0
Detection and characterization of H5N1 HPAIV in environmental samples from a dairy farm. 在一个奶牛场的环境样本中检测到 H5N1 高致病性禽流感病毒并确定其特征。
IF 1.9 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-10-01 Epub Date: 2024-07-15 DOI: 10.1007/s11262-024-02085-4
Gagandeep Singh, Jessie D Trujillo, Chester D McDowell, Franco Matias-Ferreyra, Sujan Kafle, Taeyong Kwon, Natasha N Gaudreault, Isaac Fitz, Lance Noll, Igor Morozov, Jamie Retallick, Juergen A Richt

The recent expansion of HPAIV H5N1 infections in terrestrial mammals in the Americas, most recently including the outbreak in dairy cattle, emphasizes the critical need for better epidemiological monitoring of zoonotic diseases. In this work, we detected, isolated, and characterized the HPAIV H5N1 from environmental swab samples collected from a dairy farm in the state of Kansas, USA. Genomic sequencing of these samples uncovered two distinctive substitutions in the PB2 (E249G) and NS1 (R21Q) genes which are rare and absent in recent 2024 isolates of H5N1 circulating in the mammalian and avian species. Additionally, approximately 1.7% of the sequence reads indicated a PB2 (E627K) substitution, commonly associated with virus adaptation to mammalian hosts. Phylogenetic analyses of the PB2 and NS genes demonstrated more genetic identity between this environmental isolate and the 2024 human isolate (A/Texas/37/2024) of H5N1. Conversely, HA and NA gene analyses revealed a closer relationship between our isolate and those found in other dairy cattle with almost 100% identity, sharing a common phylogenetic subtree. These findings underscore the rapid evolutionary progression of HPAIV H5N1 among dairy cattle and reinforces the need for more epidemiological monitoring which can be done using environmental sampling.

最近,美洲陆生哺乳动物感染高致病性禽流感病毒 H5N1 的范围不断扩大,包括最近在奶牛中爆发的疫情,这凸显了对人畜共患疾病进行更好的流行病学监测的迫切需要。在这项工作中,我们从美国堪萨斯州一家奶牛场采集的环境拭子样本中检测、分离并鉴定了 HPAIV H5N1。对这些样本的基因组测序发现了 PB2(E249G)和 NS1(R21Q)基因中的两个独特的置换,这在最近 2024 年流行于哺乳动物和禽类的 H5N1 病毒分离株中是罕见和不存在的。此外,约 1.7% 的序列读数显示存在 PB2(E627K)替代,这通常与病毒对哺乳动物宿主的适应有关。PB2 和 NS 基因的系统发育分析表明,该环境分离株与 2024 人感染的 H5N1 病毒分离株(A/Texas/37/2024)之间存在更多的遗传一致性。相反,HA 和 NA 基因分析表明,我们的分离物与其他奶牛中发现的分离物之间的关系更为密切,几乎 100%相同,共享一个共同的系统发育子树。这些发现凸显了高致病性禽流感病毒 H5N1 在奶牛中的快速进化过程,并加强了利用环境采样进行更多流行病学监测的必要性。
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引用次数: 0
Complete genome sequences of two tombusvirus-like viruses identified in Echinacea purpurea seeds. 在紫锥菊种子中发现的两种类墓病毒的完整基因组序列。
IF 1.9 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-10-01 Epub Date: 2024-07-18 DOI: 10.1007/s11262-024-02092-5
Juhyun Kim, Eun Jin Jeon, Minji Jun, Da-Som Lee, Seong-Jin Lee, Seungmo Lim

Echinacea is an herbaceous plant originating from North America that is cultivated for gardening and landscaping because of its showy flowers. Using high-throughput sequencing, we identified two viral contigs from echinacea seeds that were related to the family Tombusviridae. These two viruses were similar to oat chlorotic stunt virus (OCSV) and other unassigned tombusviruses; therefore, we tentatively named them Echinacea-associated tombusviruses 1 and 2 (EaTV1 and EaTV2, respectively). The EaTVs represent putative readthrough sites and have no poly(A) tails, aligning with the common features of family Tombusviridae. The EaTVs are included in a monophyletic group of OCSV and several unassigned tombusviruses. Because OCSV is the only member of Avenavirus to date, EaTVs are tentative members of Avenavirus, or they are close sister species to OCSV with several unassigned tombusviruses. RNA-dependent RNA polymerases and coat proteins were well conserved among EaTVs and unassigned tombusviruses; however, their similarities were not correlated, implying divergent and complex evolution.

紫锥花是一种原产于北美洲的草本植物,因其花朵艳丽而被栽培用于园艺和美化环境。利用高通量测序技术,我们从紫锥菊种子中鉴定出两种与Tombusviridae科相关的病毒等位基因。这两种病毒与燕麦叶枯病病毒(OCSV)和其他未确定的假花叶病毒相似;因此,我们暂时将它们命名为紫锥菊相关假花叶病毒 1 和 2(分别为 EaTV1 和 EaTV2)。EaTVs 代表推定的读穿位点,没有多聚(A)尾,符合墓病毒科的共同特征。EaTVs 包含在由 OCSV 和几种未分类的 Tombusviruses 组成的单系群中。由于 OCSV 是迄今为止 Avenavirus 的唯一成员,因此 EaTVs 是 Avenavirus 的暂定成员,或者它们是 OCSV 与几种未分类的 tombusviruses 的近亲。RNA依赖性RNA聚合酶和衣壳蛋白在EaTVs和未确定的古墓病毒之间有很好的保守性;但是,它们之间的相似性并不相关,这意味着存在着复杂的进化分歧。
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引用次数: 0
The complete genome of equid herpesvirus-1 (EHV-1) field isolates from Argentina reveals an interspecific recombinant strain. 阿根廷马疱疹病毒-1(EHV-1)野外分离株的完整基因组揭示了一个种间重组株。
IF 1.9 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-10-01 Epub Date: 2024-07-19 DOI: 10.1007/s11262-024-02093-4
Rocio Lucia Tau, Ana Eugenia Marandino, Yanina Panzera, Florencia Alamos, Maria Aldana Vissani, Sonia Alejandra Romera, Ruben Pérez, Silvina Soledad Maidana

The Equid alphaherpesvirus type 1 (EHV-1) infection can have devastating economic consequences in the horse industry due to large-scale outbreaks of abortions, perinatal foal mortality, and myeloencephalopathy. The present study analyzed the genome of two isolates obtained from aborted fetuses in Argentina, E/745/99 and E/1297/07. The E745/99 genome shares 98.2% sequence identity with Ab4, a reference EHV-1 strain. The E/1297/07 genome shares 99.8% identity with NY03, a recombinant strain containing part of ORF64 and part of the intergenic region from Equid alphaherpesvirus-4 (EHV-4). The E/1297/07 genome has the same breakpoints as other United States and Japanese recombinants, including NY03. The recombinant regions have varying numbers of tandem repeat sequences and different minor parental sequences (EHV-4), suggesting distinct origins of the recombinant events. These are the first complete genomes of EHV-1 from Argentina and South America available in the Databases.

马甲型疱疹病毒 1 型(EHV-1)感染会导致大规模的流产、围产期马驹死亡和髓脑病爆发,给养马业带来毁灭性的经济后果。本研究分析了从阿根廷流产胎儿中获得的两个分离株(E/745/99 和 E/1297/07)的基因组。E745/99 基因组与参考 EHV-1 株 Ab4 有 98.2% 的序列相同性。E/1297/07基因组与NY03有99.8%的同一性,NY03是一种重组毒株,含有ORF64的一部分和EHV-4(Equid alphaherpesvirus-4)基因间区的一部分。E/1297/07 基因组与包括 NY03 在内的其他美国和日本重组株具有相同的断裂点。重组区域具有不同数量的串联重复序列和不同的次要亲本序列(EHV-4),表明重组事件的起源不同。这些是数据库中首个来自阿根廷和南美洲的 EHV-1 完整基因组。
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引用次数: 0
Characterization of an envelope protein 118L in invertebrate iridescent virus 6 (IIV6). 无脊椎动物虹彩病毒 6(IIV6)包膜蛋白 118L 的特征。
IF 1.9 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-10-01 Epub Date: 2024-06-26 DOI: 10.1007/s11262-024-02082-7
Betul Altun, Kubra Zengin, Sevde Yayli Dabag, Aydin Yesilyurt, Remziye Nalcacioglu, Zihni Demirbag

Invertebrate iridescent virus 6 (IIV6) is a nucleocytoplasmic insect virus and a member of the family Iridoviridae. The IIV6 genome consists of 212,482 bp of linear dsDNA with 215 non-overlapping and putative protein-encoding ORFs. The IIV6 118L ORF is conserved in all sequenced members of the Iridoviridae and encodes a 515 amino acid protein with three predicted transmembrane domains and several N-glycosylation/N-myristoylation sites. In this study, we characterized the 118L ORF by both deleting it from the viral genome and silencing its expression with dsRNA in infected insect cells. The homologous recombination method was used to replace 118L ORF with the green fluorescent protein (gfp) gene. Virus mutants in which the 118L gene sequence had been replaced with gfp were identified by fluorescence microscopy but could not be propagated separately from the wild-type virus in insect cells. Unsuccessful attempts to isolate the mutant virus with the 118L gene deletion suggested that the protein is essential for virus replication. To support this result, we used dsRNA to target the 118L gene and showed that treatment resulted in a 99% reduction in virus titer. Subsequently, we demonstrated that 118L-specific antibodies produced against the 118L protein expressed in the baculovirus vector system were able to neutralize the virus infection. All these results indicate that 118L is a viral envelope protein that is required for the initiation of virus replication.

无脊椎动物虹彩病毒 6(IIV6)是一种核细胞质昆虫病毒,属于虹彩病毒科。IIV6 基因组由 212,482 bp 的线性 dsDNA 组成,其中有 215 个不重叠的假定蛋白编码 ORF。IIV6 118L ORF 在所有已测序的ridoviridae成员中都是保守的,它编码一种 515 氨基酸的蛋白质,具有三个预测的跨膜结构域和几个 N-糖基化/N-肉豆蔻酰化位点。在本研究中,我们通过从病毒基因组中删除 118L ORF 和用 dsRNA 在受感染的昆虫细胞中沉默其表达来鉴定 118L ORF 的特征。用同源重组法将 118L ORF 替换为绿色荧光蛋白(gfp)基因。用荧光显微镜鉴定了用 gfp 基因替换了 118L 基因序列的病毒突变体,但无法在昆虫细胞中与野生型病毒分开繁殖。分离 118L 基因缺失突变体病毒的尝试未获成功,这表明该蛋白对病毒复制至关重要。为了支持这一结果,我们使用 dsRNA 靶向 118L 基因,结果表明处理后病毒滴度降低了 99%。随后,我们证明了针对在杆状病毒载体系统中表达的 118L 蛋白产生的 118L 特异性抗体能够中和病毒感染。所有这些结果表明,118L 是一种病毒包膜蛋白,是病毒复制启动所必需的。
{"title":"Characterization of an envelope protein 118L in invertebrate iridescent virus 6 (IIV6).","authors":"Betul Altun, Kubra Zengin, Sevde Yayli Dabag, Aydin Yesilyurt, Remziye Nalcacioglu, Zihni Demirbag","doi":"10.1007/s11262-024-02082-7","DOIUrl":"10.1007/s11262-024-02082-7","url":null,"abstract":"<p><p>Invertebrate iridescent virus 6 (IIV6) is a nucleocytoplasmic insect virus and a member of the family Iridoviridae. The IIV6 genome consists of 212,482 bp of linear dsDNA with 215 non-overlapping and putative protein-encoding ORFs. The IIV6 118L ORF is conserved in all sequenced members of the Iridoviridae and encodes a 515 amino acid protein with three predicted transmembrane domains and several N-glycosylation/N-myristoylation sites. In this study, we characterized the 118L ORF by both deleting it from the viral genome and silencing its expression with dsRNA in infected insect cells. The homologous recombination method was used to replace 118L ORF with the green fluorescent protein (gfp) gene. Virus mutants in which the 118L gene sequence had been replaced with gfp were identified by fluorescence microscopy but could not be propagated separately from the wild-type virus in insect cells. Unsuccessful attempts to isolate the mutant virus with the 118L gene deletion suggested that the protein is essential for virus replication. To support this result, we used dsRNA to target the 118L gene and showed that treatment resulted in a 99% reduction in virus titer. Subsequently, we demonstrated that 118L-specific antibodies produced against the 118L protein expressed in the baculovirus vector system were able to neutralize the virus infection. All these results indicate that 118L is a viral envelope protein that is required for the initiation of virus replication.</p>","PeriodicalId":51212,"journal":{"name":"Virus Genes","volume":" ","pages":"549-558"},"PeriodicalIF":1.9,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141452128","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Small RNA sequencing analysis reveals regulation of microRNA expression in Madin-Darby canine kidney epithelial cells infected with Canid alphaherpesvirus 1. 小核糖核酸测序分析揭示了感染犬α疱疹病毒 1 的马丹达比犬肾上皮细胞中微小核糖核酸表达的调控。
IF 1.9 4区 医学 Q3 GENETICS & HEREDITY Pub Date : 2024-10-01 Epub Date: 2024-07-17 DOI: 10.1007/s11262-024-02091-6
Maha Ben Hamouda, Angela Pearson

Canid alphaherpesvirus 1 (CHV-1) infection can cause spontaneous abortions in pregnant dams, and in young puppies, fatal systemic infections are common. MicroRNAs (miRNAs) affect viral infection by binding to messenger RNAs, and inhibiting expression of host and/or viral genes. We conducted deep sequencing of small RNAs in CHV-1-infected and mock-infected Madin-Darby Canine Kidney (MDCK) epithelial cells, and detected sequences corresponding to 282 cellular miRNAs. Of these, 18 were significantly upregulated at 12 h post-infection, most of which were encoded on the X chromosome. We next quantified the mature forms of several of the miRNAs using stem loop RT-qPCR. Our results revealed a discordance between the levels of small RNAs corresponding to canine miRNAs, and levels of the corresponding mature miRNAs, which suggests a block in miRNA biogenesis in infected cells. Nevertheless, we identified several mature miRNAs that exhibited a statistically significant increase upon infection. These included cfa-miR-8908b, a miRNA of unknown function, and cfa-miR-146a, homologs of which target innate immune pathways and are known to play a role in other viral infections. Interestingly, ontology analysis predicted that cfa-miR-8908b targets factors involved in the ubiquitin-like protein conjugation pathway and peroxisome biogenesis among other cellular functions. This is the first study to evaluate changes in miRNA levels upon CHV-1 infection. Based on our findings, we developed a model whereby CHV-1 infection results in changes in levels of a limited number of cellular miRNAs that target elements of the host immune response, which may provide clues regarding novel therapeutic targets.

犬甲型疱疹病毒 1(CHV-1)感染可导致怀孕母犬自然流产,幼犬感染致命的全身性感染也很常见。微RNA(miRNA)通过与信使RNA结合,抑制宿主和/或病毒基因的表达,从而影响病毒感染。我们对感染 CHV-1 和模拟感染 Madin-Darby 犬肾 (MDCK) 上皮细胞中的小 RNA 进行了深度测序,检测到 282 个细胞 miRNA 的相应序列。其中有 18 种 miRNA 在感染后 12 小时明显上调,它们大多在 X 染色体上编码。接下来,我们利用干环 RT-qPCR 对几种 miRNA 的成熟形式进行了量化。我们的结果显示,与犬 miRNA 相对应的小 RNA 水平与相应的成熟 miRNA 水平不一致,这表明感染细胞中的 miRNA 生物发生受阻。尽管如此,我们还是发现了几种成熟的 miRNA 在感染后出现了统计学意义上的显著增加。其中包括功能未知的 miRNA cfa-miR-8908b 和 cfa-miR-146a,它们的同源物靶向先天性免疫通路,已知在其他病毒感染中发挥作用。有趣的是,本体分析预测,cfa-miR-8908b 以参与泛素样蛋白共轭途径和过氧物酶体生物生成等细胞功能的因子为靶标。这是第一项评估感染 CHV-1 后 miRNA 水平变化的研究。根据我们的研究结果,我们建立了一个模型,在这个模型中,CHV-1 感染会导致以宿主免疫反应要素为靶点的数量有限的细胞 miRNA 水平发生变化,这可能会为新的治疗靶点提供线索。
{"title":"Small RNA sequencing analysis reveals regulation of microRNA expression in Madin-Darby canine kidney epithelial cells infected with Canid alphaherpesvirus 1.","authors":"Maha Ben Hamouda, Angela Pearson","doi":"10.1007/s11262-024-02091-6","DOIUrl":"10.1007/s11262-024-02091-6","url":null,"abstract":"<p><p>Canid alphaherpesvirus 1 (CHV-1) infection can cause spontaneous abortions in pregnant dams, and in young puppies, fatal systemic infections are common. MicroRNAs (miRNAs) affect viral infection by binding to messenger RNAs, and inhibiting expression of host and/or viral genes. We conducted deep sequencing of small RNAs in CHV-1-infected and mock-infected Madin-Darby Canine Kidney (MDCK) epithelial cells, and detected sequences corresponding to 282 cellular miRNAs. Of these, 18 were significantly upregulated at 12 h post-infection, most of which were encoded on the X chromosome. We next quantified the mature forms of several of the miRNAs using stem loop RT-qPCR. Our results revealed a discordance between the levels of small RNAs corresponding to canine miRNAs, and levels of the corresponding mature miRNAs, which suggests a block in miRNA biogenesis in infected cells. Nevertheless, we identified several mature miRNAs that exhibited a statistically significant increase upon infection. These included cfa-miR-8908b, a miRNA of unknown function, and cfa-miR-146a, homologs of which target innate immune pathways and are known to play a role in other viral infections. Interestingly, ontology analysis predicted that cfa-miR-8908b targets factors involved in the ubiquitin-like protein conjugation pathway and peroxisome biogenesis among other cellular functions. This is the first study to evaluate changes in miRNA levels upon CHV-1 infection. Based on our findings, we developed a model whereby CHV-1 infection results in changes in levels of a limited number of cellular miRNAs that target elements of the host immune response, which may provide clues regarding novel therapeutic targets.</p>","PeriodicalId":51212,"journal":{"name":"Virus Genes","volume":" ","pages":"537-548"},"PeriodicalIF":1.9,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141629285","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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Virus Genes
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