本文選題：病毒性腦炎 + 腸道病毒��； 參考：《吉林大學(xué)》2016年碩士論文

【摘要】：背景與目的:病毒性腦炎是小兒中樞神經(jīng)系統(tǒng)感染的常見病、多發(fā)病,是危害小兒健康、致殘甚至致死的重要原因之一。引起病毒性腦炎的病原復(fù)雜多樣,主要有腸道病毒、皰疹病毒、EB病毒、腺病毒、呼吸道合胞病毒等,其中80%為腸道病毒,腸道病毒EV71型、柯薩奇病毒及�？刹《臼潜容^常見的。由于感染病毒的量不同,病毒侵犯的部位和范圍不同,臨床表現(xiàn)存在明顯差異,病情輕重不一,表現(xiàn)形式多樣,明確病原可針對性的治療。我們從2015年夏天吉林省長春市兒童醫(yī)院神經(jīng)內(nèi)科收治的病毒性腦炎確診病例中,隨機(jī)抽取95例,對患兒的腦脊液,應(yīng)用RT-PCR技術(shù)進(jìn)行病原學(xué)檢測,明確2015年夏季吉林省長春市兒童醫(yī)院收治病毒性腦炎病原;同時回顧性分析患兒臨床特點。臨床特點與病原學(xué)診斷方法的研究,對于早期診斷、有效治療有重要意義。探討應(yīng)用簡并引物PCR擴(kuò)增產(chǎn)物直接測序技術(shù)在小兒腸道病毒性腦炎診斷中的應(yīng)用價值,為病毒性腦炎的病原學(xué)診治提供理論依據(jù)。方法:研究對象:2015年夏天吉林省長春市兒童醫(yī)院神經(jīng)內(nèi)科確診的病毒性腦炎患兒95例;研究方法:首先,應(yīng)用實時熒光定量PCR方法對患兒腦脊液標(biāo)本進(jìn)行腸道病毒通用型、柯薩奇病毒A6型和A10型、EB病毒核酸檢測;然后,再從腸道病毒通用型陽性標(biāo)本中抽取40例標(biāo)本,應(yīng)用簡并引物PCR擴(kuò)增;最后,從中選取20例電泳條帶比較理想的擴(kuò)增產(chǎn)物進(jìn)行基因測序,測序結(jié)果在NCBI上應(yīng)用BLAST在線核酸序列分析,確定腸道病毒型別。同時回顧性分析患兒的一般情況及臨床特點。結(jié)果:1.95例患兒腸道通用型實時熒光定量PCR檢測陽性為84例(88.4%),EB病毒PCR檢測3例陽性(3.15%),柯薩奇病毒A6型和A10型均為陰性。2.84例腸道病毒通用型陽性患兒臨床特點為:發(fā)熱、頭痛、嘔吐,其它系統(tǒng)癥狀少見,體征主要為巴氏征陽性,少部分伴有項強(qiáng)陽性,預(yù)后良好。3.20份簡并引物擴(kuò)增產(chǎn)物,且電泳條帶清晰的標(biāo)本進(jìn)行序列測定及基因比對,結(jié)果有14例為Echo30型腸道病毒,其余6例未找到匹配結(jié)果。結(jié)論:2015年夏天吉林省局部地區(qū)病毒性腦炎流行病原為Echo30型腸道病毒,該病毒性腦炎,以6-9月份為流行高峰,傳染性強(qiáng),集中發(fā)病,臨床表現(xiàn)主要癥狀為發(fā)熱、頭痛、嘔吐,其它系統(tǒng)癥狀少見,體征主要為巴氏征陽性,少部分伴有項強(qiáng)陽性,預(yù)后良好。簡并引物RT-PCR擴(kuò)增物可進(jìn)行基因測序,快速明確病原。因同一病原可引起不同的臨床表現(xiàn),而相同的臨床表現(xiàn)亦可由不同病原引起,故明確病原后歸納其臨床特點,對今后病毒性腦炎的預(yù)防及疫苗的研制提供有效的參考依據(jù)。
[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.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R725.1

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