盘锦地区呼吸道感染病毒病原检测及分离

发布时间：2018-03-10 21:33

本文选题：急性呼吸道感染　切入点：实时荧光定量PCR　出处：《贵州师范大学》2017年硕士论文　论文类型：学位论文

【摘要】：急性呼吸道感染(Acute Respiratory Infections,ARI)是急性感染性疾病及其死亡病例的主要原因,是全球范围内的公共卫生问题。ARI可由多种病原微生物引起,其中,病毒是引起ARI的主要病原体。因此,开展ARI病原检测和分离,对于ARI流行趋势的监控以及预防、控制都具有重要的现实意义。本研究以2013年4月-2014年3月期间盘锦地区ARI患者咽拭子样品作为实验材料,进行病毒病原检测,分析其构成,对新发传染病流行征兆、重大传染源、早期病例等进行预判,以避免突发公共卫生事件的发生。研究结果如下:1、采用核酸提取、荧光定量PCR方法,对ARI患者咽拭子样品856份进行流感病毒(H1N1、H3N2、IVB)、合胞病毒(RSV)、副流感病毒(PIV1、PIV2、PIV3)、腺病毒(AdV)、偏肺病毒(MPV)、鼻病毒(HRV)检测,表明:(1)流感病毒、副流感病毒、鼻病毒和腺病毒是本地区引起ARI的主要病毒病原体,其阳性样品检出率分别为72.38%、5.71%、6.67%和10.48%;(2)在流感病毒阳性样品中,检出H1N1型67株、H3N2型9株,分别占阳性样品检出率的88.16%和11.84%,但未检出乙型流感病毒流感病毒;2、在2013-2014年,盘锦地区流感病毒的流行存在以下规律:冬季为高发期,夏季最低;发病年龄主要集中在5-14岁年龄组,而25-59岁组最低;以散发病例为主,未见流感爆发疫情,其中,新甲型H1为优势流行株,H3N2亚型少量出现。3、通过对盘锦地区流感病毒的MDCK细胞病毒分离和鸡胚病毒分离比较,发现流感病毒对鸡胚的敏感性仍然弱于MDCK细胞分离,但是在流感病毒疫苗生产中不能直接使用MDCK细胞分离病毒,因此需要将一次MDCK细胞分离和鸡胚分离要时进行,由此分离出的毒株可送国家流感中心用于试剂的制备和疫苗生产。综上所述,通过对盘锦地区2013年4月-2014年3月期间呼吸道感染病毒病原进行采样,利用分子生物学检测方法对病原微生物进行检测及分离,初步掌握本地区呼吸道感染病例样本中主要病毒种类及发病规律,并建立了MDCK细胞分离和鸡胚分离合并使用的流感病毒分离方法。本文研究结果为临床上医师用药、疫苗研制、突发公共卫生事件预警、指挥决策和现场处理提供一定科学依据。
[Abstract]:Acute Respiratory infection (ARI) is a major cause of acute infectious diseases and death cases, and a global public health problem. ARI can be caused by a variety of pathogenic microorganisms, among which viruses are the main pathogens of ARI. The detection and separation of ARI pathogens are of great practical significance for the monitoring, prevention and control of ARI epidemic trend. The pharynx swab samples of ARI patients in Panjin region from April 2013 to March 2014 were used as experimental materials in this study. Virus pathogen detection, analysis of its composition, predetermination of epidemic signs of new infectious diseases, major sources of infection, early cases, etc., in order to avoid the occurrence of sudden public health incidents. The research results are as follows: 1, extracted by nucleic acid. Fluorescence quantitative PCR was used to detect 856 throat swabs from ARI patients with influenza virus H1N1H3N2OIVBV, syncytial virus (RSVV), parainfluenza virus (PIV1) PIV2PIV3, adenovirus (AdVo), parapneumovirus (MPV1), rhinovirus (rhinovirus HRV), which showed that influenza virus, parainfluenza virus, parainfluenza virus, parainfluenza virus, parainfluenza virus, parainfluenza virus, parainfluenza virus, parainfluenza virus, and parainfluenza virus. Rhinovirus and adenovirus are the main pathogens causing ARI in this area. The positive rates of rhinovirus and adenovirus are 72.38% and 10.48% respectively. The positive rate of influenza virus was 88.16% and 11.84, respectively, but no influenza B virus was detected. In 2013-2014, the epidemic pattern of influenza virus in Panjin area was as follows: high incidence in winter and lowest in summer; The age of onset was mainly in the 5-14 age group, while the 25-59 age group was the lowest. New A / H _ 1 was the dominant epidemic strain. A few subtypes of H3N2 appeared in Panjin area. By comparing the isolation of MDCK cell virus and chicken embryo virus from Panjin area, it was found that the sensitivity of influenza virus to chicken embryo was still weaker than that of MDCK cell isolation. But MDCK cells cannot be used directly to isolate the virus in flu vaccine production, so it is necessary to separate MDCK cells and chicken embryos at the same time. The strains isolated from the virus can be sent to the National Influenza Center for preparation of reagents and vaccine production. To sum up, by sampling the pathogens of respiratory tract infections from April 2013 to March 2014 in Panjin region, Molecular biological methods were used to detect and isolate pathogenic microbes in order to master the main virus types and pathogenesis patterns of respiratory tract infection cases in this area. The method of influenza virus isolation used in combination of MDCK cell isolation and chicken embryo isolation was established. The results provide scientific basis for clinical medicine, vaccine development, public health emergency warning, command decision-making and field treatment.
【学位授予单位】：贵州师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R56

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