本文選題：急性呼吸道感染　切入點(diǎn)：實(shí)時(shí)熒光定量PCR　出處：《貴州師范大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【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,是全球范圍內(nèi)的公共衛(wèi)生問(wèn)題。ARI可由多種病原微生物引起,其中,病毒是引起ARI的主要病原體。因此,開(kāi)展ARI病原檢測(cè)和分離,對(duì)于ARI流行趨勢(shì)的監(jiān)控以及預(yù)防、控制都具有重要的現(xiàn)實(shí)意義。本研究以2013年4月-2014年3月期間盤錦地區(qū)ARI患者咽拭子樣品作為實(shí)驗(yàn)材料,進(jìn)行病毒病原檢測(cè),分析其構(gòu)成,對(duì)新發(fā)傳染病流行征兆、重大傳染源、早期病例等進(jìn)行預(yù)判,以避免突發(fā)公共衛(wèi)生事件的發(fā)生。研究結(jié)果如下:1、采用核酸提取、熒光定量PCR方法,對(duì)ARI患者咽拭子樣品856份進(jìn)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)檢測(cè),表明:(1)流感病毒、副流感病毒、鼻病毒和腺病毒是本地區(qū)引起ARI的主要病毒病原體,其陽(yáng)性樣品檢出率分別為72.38%、5.71%、6.67%和10.48%;(2)在流感病毒陽(yáng)性樣品中,檢出H1N1型67株、H3N2型9株,分別占陽(yáng)性樣品檢出率的88.16%和11.84%,但未檢出乙型流感病毒流感病毒;2、在2013-2014年,盤錦地區(qū)流感病毒的流行存在以下規(guī)律:冬季為高發(fā)期,夏季最低;發(fā)病年齡主要集中在5-14歲年齡組,而25-59歲組最低;以散發(fā)病例為主,未見(jiàn)流感爆發(fā)疫情,其中,新甲型H1為優(yōu)勢(shì)流行株,H3N2亞型少量出現(xiàn)。3、通過(guò)對(duì)盤錦地區(qū)流感病毒的MDCK細(xì)胞病毒分離和雞胚病毒分離比較,發(fā)現(xiàn)流感病毒對(duì)雞胚的敏感性仍然弱于MDCK細(xì)胞分離,但是在流感病毒疫苗生產(chǎn)中不能直接使用MDCK細(xì)胞分離病毒,因此需要將一次MDCK細(xì)胞分離和雞胚分離要時(shí)進(jìn)行,由此分離出的毒株可送國(guó)家流感中心用于試劑的制備和疫苗生產(chǎn)。綜上所述,通過(guò)對(duì)盤錦地區(qū)2013年4月-2014年3月期間呼吸道感染病毒病原進(jìn)行采樣,利用分子生物學(xué)檢測(cè)方法對(duì)病原微生物進(jìn)行檢測(cè)及分離,初步掌握本地區(qū)呼吸道感染病例樣本中主要病毒種類及發(fā)病規(guī)律,并建立了MDCK細(xì)胞分離和雞胚分離合并使用的流感病毒分離方法。本文研究結(jié)果為臨床上醫(yī)師用藥、疫苗研制、突發(fā)公共衛(wèi)生事件預(yù)警、指揮決策和現(xiàn)場(chǎng)處理提供一定科學(xué)依據(jù)。
[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.
【學(xué)位授予單位】：貴州師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R56

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本文編號(hào)：1595158