2015年吉林省部分地区95例病毒性脑炎临床特点及病原学检测

发布时间：2018-04-23 03:22

本文选题：病毒性脑炎 + 肠道病毒　；参考：《吉林大学》2016年硕士论文

【摘要】：背景与目的:病毒性脑炎是小儿中枢神经系统感染的常见病、多发病,是危害小儿健康、致残甚至致死的重要原因之一。引起病毒性脑炎的病原复杂多样,主要有肠道病毒、疱疹病毒、EB病毒、腺病毒、呼吸道合胞病毒等,其中80%为肠道病毒,肠道病毒EV71型、柯萨奇病毒及埃可病毒是比较常见的。由于感染病毒的量不同,病毒侵犯的部位和范围不同,临床表现存在明显差异,病情轻重不一,表现形式多样,明确病原可针对性的治疗。我们从2015年夏天吉林省长春市儿童医院神经内科收治的病毒性脑炎确诊病例中,随机抽取95例,对患儿的脑脊液,应用RT-PCR技术进行病原学检测,明确2015年夏季吉林省长春市儿童医院收治病毒性脑炎病原;同时回顾性分析患儿临床特点。临床特点与病原学诊断方法的研究,对于早期诊断、有效治疗有重要意义。探讨应用简并引物PCR扩增产物直接测序技术在小儿肠道病毒性脑炎诊断中的应用价值,为病毒性脑炎的病原学诊治提供理论依据。方法:研究对象:2015年夏天吉林省长春市儿童医院神经内科确诊的病毒性脑炎患儿95例;研究方法:首先,应用实时荧光定量PCR方法对患儿脑脊液标本进行肠道病毒通用型、柯萨奇病毒A6型和A10型、EB病毒核酸检测;然后,再从肠道病毒通用型阳性标本中抽取40例标本,应用简并引物PCR扩增;最后,从中选取20例电泳条带比较理想的扩增产物进行基因测序,测序结果在NCBI上应用BLAST在线核酸序列分析,确定肠道病毒型别。同时回顾性分析患儿的一般情况及临床特点。结果:1.95例患儿肠道通用型实时荧光定量PCR检测阳性为84例(88.4%),EB病毒PCR检测3例阳性(3.15%),柯萨奇病毒A6型和A10型均为阴性。2.84例肠道病毒通用型阳性患儿临床特点为:发热、头痛、呕吐,其它系统症状少见,体征主要为巴氏征阳性,少部分伴有项强阳性,预后良好。3.20份简并引物扩增产物,且电泳条带清晰的标本进行序列测定及基因比对,结果有14例为Echo30型肠道病毒,其余6例未找到匹配结果。结论:2015年夏天吉林省局部地区病毒性脑炎流行病原为Echo30型肠道病毒,该病毒性脑炎,以6-9月份为流行高峰,传染性强,集中发病,临床表现主要症状为发热、头痛、呕吐,其它系统症状少见,体征主要为巴氏征阳性,少部分伴有项强阳性,预后良好。简并引物RT-PCR扩增物可进行基因测序,快速明确病原。因同一病原可引起不同的临床表现,而相同的临床表现亦可由不同病原引起,故明确病原后归纳其临床特点,对今后病毒性脑炎的预防及疫苗的研制提供有效的参考依据。
[Abstract]:Background & objective: viral encephalitis is a common disease of central nervous system infection in children. The pathogens of viral encephalitis are complex and diverse, including enterovirus, herpesvirus Epstein-Barr virus, adenovirus, respiratory syncytial virus and so on. 80% of them are enterovirus, enterovirus EV71 type, coxsackie virus and Echo virus are more common. Because the quantity of infected virus is different, the site and scope of virus invasion are different, the clinical manifestation is obviously different, the severity of disease is different, the form of expression is various, and it is clear that the pathogen can be targeted for treatment. We randomly selected 95 cases of viral encephalitis from the neurology department of Changchun Children's Hospital of Jilin Province in the summer of 2015. The cerebrospinal fluid (CSF) of the children was detected by RT-PCR technique. To identify the pathogen of viral encephalitis in Changchun Children's Hospital of Jilin Province in summer of 2015, and to analyze retrospectively the clinical characteristics of children with viral encephalitis. The study of clinical characteristics and etiological diagnosis is of great significance for early diagnosis and effective treatment. To explore the value of direct sequencing of degenerate primer PCR amplification products in the diagnosis of viral encephalitis in children and to provide theoretical basis for etiological diagnosis and treatment of viral encephalitis. Methods: 95 children with viral encephalitis diagnosed in neurology department of Changchun Children's Hospital of Jilin Province in summer of 2015 were studied. Coxsackie virus A6 and A10 Epstein-Barr virus nucleic acid detection; then, from the enterovirus common positive specimens, 40 samples, using degenerate primers PCR amplification; finally, Among them, 20 cases were selected for gene sequencing, and BLAST on-line nucleic acid sequence analysis was used to determine the type of enterovirus on NCBI. At the same time, the general situation and clinical characteristics of children were analyzed retrospectively. Results the positive rate of real-time fluorescent quantitative PCR was 88. 4% in 84 children with EBV PCR, 3 cases were positive for EBV PCR. The clinical characteristics of Coxsackie virus A6 and A10 were negative. 2.84 cases of common type positive of enterovirus were fever and headache. Vomiting, other systemic symptoms were rare, the signs were mainly Pap sign positive, a few with strong positive items, good prognosis of 3.20 degenerate primer amplification products, and electrophoretic bands of clear samples for sequencing and gene alignment, Results there were 14 cases of Echo30 enterovirus, the other 6 cases did not find a matched result. Conclusion: the epidemic of viral encephalitis in local area of Jilin Province in summer of 2015 was originally Echo30 enterovirus. The epidemic peak of viral encephalitis was June to September, which was highly infectious and concentrated. The main clinical symptoms were fever, headache and vomiting. Other systemic symptoms were rare, the signs were mainly Pap sign positive, a few accompanied with strong positive item, the prognosis was good. Degenerate primer RT-PCR amplifiers can be sequenced to identify the pathogen quickly. Since the same pathogen can cause different clinical manifestations and the same clinical manifestations can also be caused by different pathogens, the clinical characteristics of the same pathogen can be summed up, which will provide an effective reference basis for the prevention of viral encephalitis and the development of vaccine in the future.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R725.1

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