Detection and genome characterization of Middelburg virus strains isolated from CSF and whole blood samples of humans with neurological manifestations in South Africa.
Detection and genome characterization of Middelburg virus strains isolated from CSF and whole blood samples of humans with neurological manifestations in South Africa.
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DOI：
10.1371/journal.pntd.0010020
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发表时间：
2022-01
影响因子：
3.8
通讯作者：
Venter M
中科院分区：
文献类型：
--
作者：
Fourie I;Williams J;Ismail A;Jansen van Vuren P;Stoltz A;Venter M
东半球甲型病毒-米德尔堡病毒(MIDV)并不为人所知，尽管之前曾在南部非洲描述过几例与动物疾病有关的病例，但尚未对该病毒与人类疾病的联系进行调查。目前的研究旨在调查MIDV感染与南非豪登省住院或有症状的患者发烧或神经症状的可能联系。本研究是一项基于实验室的描述性、回溯性和前瞻性研究。从豪登州公立医院提交给国家卫生实验室服务(NHLS)的Tshwane学术部门用于病毒研究的存档脑脊液(CSF)样本，以及来自患有神经系统和发热性疾病的兽医学生特别病例的EDTA(乙二胺四乙酸乙酯)全血样本，被选择并使用实时逆转录(RtRT)PCR法筛查甲型病毒的存在。从rtRT-PCR阳性样本中分离的病毒在Vero细胞培养中进行，并用于获得全基因组序列。使用EpiInfo进行基本描述性统计分析。从豪登州茨瓦内地区住院患者的NHL中获得了3/187份回顾性脑脊液样本，并从来自比勒陀利亚兽医大学兽医系学生的特别查询虫媒病毒病例中提取了1/2 EDTA样本，通过rtRT-PCR检测了MIDV。从两例病例中获得了病毒分离株的全基因组序列，其中一例来自EDTA全血样本(特别病例)，另一例来自脑脊液样本(NHLS样本)。在四例Middelburg病毒阳性病例中，有两例在感染时有其他合并症或感染。对有神经症状的患者脑脊液中MIDV的检测表明，应将该病毒作为一种人类病原体进行调查，该病毒可能会引起或促成儿童和成人的神经症状。旧大陆甲型病毒主要与人类多发性关节炎有关。基孔肯雅病毒在全球范围内的出现引起了人们对被忽视的非洲虫媒病毒的潜在疾病的关注。新的世界甲型病毒与美洲人和马的严重神经系统疾病有关，死亡率很高。米德尔堡病毒(MIDV)是一种东半球甲型病毒，以前与南部非洲马匹和野生动物的神经系统疾病和死亡有关，但尚未与人类疾病有关。这项研究调查了南非出现急性神经系统症状的人中的MIDV。使用普通甲型病毒实时RT-PCR和系统发育分析，在189例此类病例中有4例(2%)确认了MIDV感染。MIDV的全基因组是从一名2岁儿童的脑脊液样本和一名兽医学生的全血样本中获得的，两人都表现出神经症状。这项研究表明，MIDV可能与非洲人类未解决的神经疾病有关。
The Old world Alphavirus, Middelburg virus (MIDV), is not well known and although a few cases associated with animal illness have previously been described from Southern Africa, there has been no investigation into the association of the virus with human illness. The current study aimed to investigate possible association of MIDV infection with febrile or neurological manifestations in hospitalized or symptomatic patients fromGauteng, South Africa. This study is a descriptive retrospective and prospective laboratory based study. Archived cerebrospinal fluid (CSF) samples submitted to the National Health Laboratory Service (NHLS), Tshwane Academic division for viral investigation from public sector hospitals in Gauteng as well as EDTA (ethylenediaminetetraacetic acid) whole blood samples from ad hoc cases of veterinary students, presenting with neurological and febrile illness, were selected and screened for the presence of alphaviruses using real-time reverse transcription(rtRT) PCR.Virus isolations from rtRT-PCR positive samples were conducted in Vero cell culture and used to obtain full genome sequences. Basic descriptive statistical analysis was conducted using EpiInfo. MIDV was detected by rtRT-PCR in 3/187 retrospective CSF specimens obtained from the NHLS from hospitalised patients in the Tshwane region of Gauteng and 1/2 EDTA samples submitted in the same year (2017) from ad hoc query arbovirus cases from veterinary students from the Faculty of Veterinary Science University of Pretoria.Full genome sequences were obtained for virus isolates from two cases; one from an EDTA whole blood sample (ad hoc case) and another from a CSF sample (NHLS sample).Two of the four Middelburg virus positive cases,for which clinical information was available, had other comorbidities or infections at the time of infection. Detection of MIDV in CSF of patients with neurological manifestations suggests that the virus should be investigated as a human pathogen with the potential of causing or contributing to neurological signs in children and adults. Old World alphaviruses are mainly associated with polyarthritis in humans. Global emergence of chikungunya virus raises concerns regarding disease potential of neglected African arboviruses.New World alphaviruses are associated with severe neurological disease in humans and horses in the Americas with a high mortality rate. Middelburg virus (MIDV) is an Old World alphavirus previously associated with neurological disease and fatalities in horses and wildlife in southern Africa but not yet linked to disease in humans. This study investigated MIDV in humans presenting with acute neurological symptoms in South Africa. MIDV infection was confirmed in 4/189 (2%) such cases using a generic Alphavirus real-time RT-PCR and phylogenetic analysis. MIDV full genomes were obtained from a cerebrospinal fluid sample of a 2-year-old child and a whole blood sample of a veterinary student, both presenting with neurological signs. This study suggests MIDV may be associated with unsolved neurological disease in humans in Africa.