569例呼吸道感染住院儿童7种常见呼吸道病毒病原学分析

目的了解呼吸道感染住院患儿呼吸道病毒的分布情况。方法选取2015年7月至2016年6月呼吸道感染的住院患儿病例,抽取鼻咽分泌物,采用直接免疫荧光法检测常见的7种病毒,即呼吸道合胞病毒(RSV)、甲型流感病毒(IVA)、乙型流感病毒(IVB)、副流感病毒1型(PIV1)、副流感病毒2型(PIV2)、副流感病毒3型(PIV3)、腺病毒(IVD)。结果共有569例样本送检,193例病毒检测阳性(33.92%),其中有3例为2种病毒的混合感染。男性共369例,女性共200例。其中RSV、PIV3、ADV的感染率居前三位。呼吸道病毒的感染率随着患儿年龄的增长逐渐下降,除新生儿外,6个月内的婴幼儿呼吸道病毒检出率最高。RSV检测阳性率自11月开始呈增高趋势,12月最高,达55.86%。PIV3的检出高峰出现在7月,达13.64%。结论 RSV是本地区儿童呼吸道感染最常见的病毒。呼吸道病毒的检出率与年龄、季节等因素有关。     

503例儿童急性呼吸道感染常见病毒的检测结果分析

目的通过对503例急性呼吸道感染患儿进行7种常见病毒[流感病毒A、B型（IVA、IVB）,腺病毒（ADV）,副流感病毒1、2、3型（PIV1、PIV2、PIV3）及呼吸道合胞病毒（RSV）]的检测,了解本地区2013年急性呼吸道感染患儿的病毒感染状况。方法应用免疫荧光法对503例急性呼吸道感染患儿的咽拭子进行7种常见病毒的检测。结果 503例患儿中有55例检出病毒阳性,总阳性率为10.93%,均为单一病毒感染。7种常见病毒中,RSV的感染率最高,为76.36%。在各年龄组中,〈1岁组的病毒检出率最高,为29.41%,随着年龄的增长,检出率逐渐降低。从季节分布来看,春季的感染率最高,为23.08%,其次为冬季,感染率10.13%。结论 RSV是2013年本地区儿童急性呼吸道感染的主要病毒,〈1岁组患儿的病毒检出率最高,春季为感染的高发季节。     

南京地区2010～2011年度呼吸道感染患儿腺病毒的流行病学研究

本研究为了解南京地区儿童腺病毒(ADV)感染的流行特点及型别,收集2010年8月至2011年7月南京医科大学附属南京儿童医院住院及门诊呼吸道感染患儿的鼻咽抽吸物(NPA)及咽拭子(NPS)共644例,采用巢氏聚合酶链反应法(Nested-PCR)检测ADV hexon基因,将阳性PCR扩增产物进行测序、同源性和进化分析。同时对12种其他呼吸道相关病毒进行PCR检测,包括人博卡病毒(HBoV),呼吸道合胞病毒(RSV),人鼻病毒(HRV),副流感病毒1～4型(PIV1-4),流感病毒A和B(IFVA/B),人偏肺病毒(HMPV),冠状病毒NL63和HKU1(HCoV-NL63和HCoV-HKU1)。结果显示:644例标本中共检出ADV阳性扩增产物171份,检出率为26.55%,3型120例(70.18%,120/171),7型16例(9.36%,16/171),1型12例(7.02%,12/171),2型10例(5.85%,10/171),5型6例(3.51%,6/171),6型3例(1.75%,3/171),57型3例(1.75%,3/171),41型1例(0.58%,1/171)。ADV感染呈全年散发,其发病高峰主要在4～7月。以7岁以下儿童多见(96.49%)。171例ADV感染患儿中有99例(57.89%)存在混合感染,其中以呼吸道合胞病毒(RSV)、人鼻病毒(HRV)多见。ADV阳性患儿诊断以下呼吸道感染为主(63.16%),肺炎占30.41%。结论:ADV是2010年8月到2011年7月南京地区儿童呼吸道感染的重要病原之一,其优势流行株为3型,长期监测其流行型别具有重要意义。     

大连地区住院患儿急性呼吸道感染流行病学调查

目的分析大连地区急性呼吸道感染患儿病原体检测结果及流行情况,为临床早期诊断、治疗及预防提供依据。方法调查2015年1月-2016年7月医院住院急性呼吸道感染患儿2 944例,采用间接免疫荧光法(IFA)对呼吸道感染患儿血液中的呼吸道合胞病毒(respiratory syncytial virus,RSV)、腺病毒(adenovirus,ADV)、流感病毒A型(influenza virus type A,IFA)、流感病毒B型(influenza virus type B,IFB)、副流感病毒(parainfluenza virus,PIV)、肺炎衣原体(Chlamydia pneumoniae,CP)、肺炎支原体(Mycoplasma pneumonia,MP)和嗜肺军团菌(Legionella pneumophila,LP)8种病原体进行检测。结果 2 944例患儿中血清IgM抗体阳性共1 280例,总阳性率为43.48%(1 280/2 944),主要为IFB和MP感染:IFB阳性率最高,为16.68%(491/2 944);其次为MP,阳性率为14.44%(425/2 944)。ADV各年龄组之间差异无统计学意义(χ~2=0.286,P=0.5930.05),其余各种病原体不同年龄组之间差异均有统计学意义。其中IFB、MP在3~6岁组中检出率最高;RSV在0~1岁组中检出率明显高于其他年龄组。MP男女患儿组间差异有统计学意义(χ~2=7.671,P=0.0060.05),其余各项差异均无统计学意义(P0.05)。IFB第二季度和第三季度检出率高于其他季度,MP在第三季度检出率最高,IFA、LP、CP也同样是第三季度检出率最高,PIV和ADV各季度检出率差异无统计学意义(P0.05),RSV在第一季度检出率略高于其他季度。结论大连地区引起患儿呼吸道感染的病原体主要是IFB和MP,针对不同年龄组、不同季度的感染情况,应做好重点预防工作。     

2009～2010年上海地区急性呼吸道感染病毒病原谱分析

调查2009~2010年上海地区人群急性呼吸道感染(ARTI)的病毒性病原,探讨2009甲型H1N1流感暴发背景下呼吸道感染病毒病原谱的构成。采用套式多重反转录-聚合酶链反应(RT-PCR)和实时荧光定量RT-PCR方法,对来自2 044例患者的2 044份标本(包括2 005份鼻咽拭子和39份肺泡灌洗液),同时检测腺病毒(ADV)、副流感病毒(PIV)、甲型流感病毒(FluA)、乙型流感病毒(FluB)、微小核糖核酸病毒、呼吸道合胞病毒(RSV)、人偏肺病毒(hMPV)、冠状病毒(CoV)和人博卡病毒(HBoV)。其中,656(32.09%)份标本经呼吸道病毒检测为阳性,52份标本为双重感染。FluA检出率最高(13.36%),其后依次为微小核糖核酸病毒(10.23%)、FluB(4.84%)、ADV(1.96%)、PIV(1.76%)、RSV(1.32%)、CoV(0.59%)、hMPV(0.39%)和HBoV(0.20%)。但各月病毒检出率分布不均,2009和2010年呼吸道病毒检出率高峰出现在当年11月(53.07%和65.59%),低谷都出现在当年5月,且2009年5~9月的病毒检出率高于2010年同期(32.02%vs15.38%,P<0.05)。其中,2009甲型H1N1流感暴发导致2009年10月~2010年1月2009甲型H1N1流感病毒占当月检出FluA的100%,2009年6~9月也占当月检出FluA的较高比率,依次为90.91%(20/22)、75.00%(15/20)、48.00%(12/25)和56.25%(18/32)。比较甲型H3N2流感病毒和2009甲型H1N1流感病毒分别在上呼吸道感染(URTI)和下呼吸道感染(LRTI)中的检出率,无统计学差异(URTI,85.29%vs76.61%;LRTI,14.71%vs23.39%;P>0.05)。呼吸道病毒检出率还与年龄相关,0~4岁组和5~14岁组病毒检出率高于其他年龄组。在0~4岁及≥65岁组中,微小核糖核酸病毒检出率最高,FluA次之;其余年龄组中FluA检出率最高。混合感染中15岁以下儿童占50%(26/52),微小核糖核酸病毒与其他病毒混合感染占84.62%(44/52)。本研究表明,上海地区2009~2010年FluA是最常见的急性呼吸道感染病原,2009甲型H1N1流感病毒成为2009年FluA的优势亚型。微小核糖核酸病毒是混合感染中最常见的病原。结果提示,应长期监测主要呼吸道病毒的活动水平,并加强对微小核糖核酸病毒流行病学和致病性的研究。     

上海复旦大学附属儿科医院急性下呼吸道感染患儿常见病毒的检测及临床研究

目的 建立套式反转录-聚合酶链反应(nested-RT-PCR)检测临床样本鼻病毒(human rhinovirus,HRV)基因的方法和直接免疫荧光(DFA)的方法对常见呼吸道病毒病原进行检测,了解上海儿科医院小儿急性下呼吸道感染(ALRTI)中病毒感染的现状.方法 收集住院确诊为ALRTI患儿的深部鼻咽分泌物(NPS)标本.用套式RT-PCR的方法检测NPS中的HRV基因,并进一步对扩增产物进行序列分析;用DFA的方法检测NPS中的呼吸道合胞病毒(RSV)、流感病毒(IFV)、副流感病毒(PIV)、腺病毒(ADV)抗原.并分析各种病毒所致ALRTI的临床特点及流行特征.结果 342例患儿除细菌感染外共130例病毒抗原阳性,其中检测出RSV 64例(18.70%),1岁以下婴儿的RSV检出数为71.8%.RSV 8月份起检出率逐渐升高,到12月份达到最高;HRV阳性患儿46例(13.45%),3岁以下婴幼儿检出例数有38例,占HRV检出数的82.6%.HRV致ALRTI全年可见,在3到5月为最高峰;ADV感染9例(2.63%);PIV感染8例(2.3%);IFV感染7例(2.0%);病毒合并感染共有4例.46例HRV阳性标本中选取15份用另1对引物扩增并测序,测得的核苷酸序列经过blast比较,与GenBank中已知HRV序列的同源性在83%～97%,样本间的序列比较提示变异性较大.130例病毒性ALRTI患儿绝大部分诊断为支气管肺炎.以中等度发热为主,末梢血白细胞＜10×109/L 93例(71.5%),CRP＜8 mg/L 99例(76.2%),符合病毒性肺炎的特点,并发症较少,均治愈出院.结论 本地区小儿病毒性ALRTI中,以RSV及HRV占为主要地位,ADV、PIV及IFV相对少见.较少有混合性病毒感染.HRV所致小儿ALRTI以婴幼儿居多,集中于春、秋两季.套式RT-PCR检测HRV基因快速、操作简便、特异性较强,HRV基因组具有高度变异的特点.     

629例住院儿童临床七种常见呼吸道病毒分布特征

目的了解本地区七种常见呼吸道病毒临床分布特征,旨在为呼吸道病毒的预防、治疗提供帮助。方法随机选取2 001例呼吸科住院患者样本,采用直接免疫荧光法进行呼吸道病毒抗原检测。结果 2 001例患者样本中,共检测出629例阳性病毒,阳性率为31.43%,阳性率前三位的病毒依次为甲型流感病毒(FluA)、呼吸道合胞病毒(RSV)、乙型流感病毒,分别为12.14%、9.90%和3.80%,而副流感病毒II型检出率最低,为0.50%。冬、春季节为呼吸道病毒高发季节,其中一月和二月的检出率高达56.91%和43.15%;而夏、秋季节检出率相对较低,六月的检出率仅为7.00%;1岁年龄组病毒阳性检出率最高,且随年龄增大,阳性检出率明显降低。结论冬、春季节是呼吸道病毒的高发季节;FluA和RSV是呼吸道感染中最常见的病毒;1岁年龄组病毒阳性检出率最高。     

冬季急性下呼吸道感染住院儿童病原学分析及临床流行病学特点

目的了解冬季儿童急性下呼吸道感染病原谱及临床流行病学特征,为临床抗感染及病原检测提供依据。方法对我院2006年12月～2007年2月急性下呼吸道感染住院患儿采用一次性无菌吸痰管经鼻腔插入7～8cm,达到咽部以下负压吸取1～2ml深部鼻咽分泌液送细菌培养,并对呼吸道合胞病毒(RSV),腺病毒(ADV),A、B型流感病毒(IFV),1、2及3型副流感病毒(PIV)等7种常见呼吸道病毒抗原进行检测及运用荧光定量聚合酶链反应(PCR)技术检测标本中支原体和衣原体DNA。结果①381份下呼吸道感染儿童痰标本中细菌培养阳性81份,病毒检测阳性133份,支原体和衣原体阳性分别12份与6份,混合感染标本44份,总标本病原学检出率为50.66%(193/381)。②RSV阳性标本112份,为最重要的感染病原,连续3个月RSV检出率均在30%左右,6月龄以下儿童占61.61%,2岁以下儿童占86.61%,阳性标本中78.57%的患儿有喘息表现。③大肠埃希菌(16株)、肺炎链球菌(14株)、肺炎克雷伯菌及金黄色葡萄球菌(各10株)、卡他莫拉菌布兰汉亚种(8株)、流感嗜血杆菌和副流感嗜血杆菌(各6株)表现为主要的致病菌。结论RSV感然仍为儿童冬季急性下呼吸道感染最主要的病原,尤其在2岁以下儿童,且易表现为喘息发作。仍有40%以上感染病原未明,儿童急性下呼吸道感染病原谱需进一步完善。     

九项呼吸道感染病原体IgM检测结果分析

目的 探讨新乡市第一人民医院呼吸系统感染患者9种呼吸道病毒的分布情况.方法 回顾性分析该院门诊和住院的呼吸系统感染者中接受病毒抗体检测330例患者的检测结果.结果 入选病例共330例,检测出抗体阳性者193例,阳性检出率为58.48％,其中肺炎支原体(MP) 131例(39.70％),嗜肺军团菌(LP) 89例(26.97％),肺炎衣原体(CPn) 17例(5.15％),Q热立克次体(COX)15例(4.55％),副流感病毒(PIVs)7例(2.12％),呼吸道合胞病毒(RSV)6例(1.82％),甲型流感病毒(INFA)5例(1.52％),乙型流感病毒(INFB)3例(0.91％),腺病毒(ADV)2例(0.61％).结论 MP、LP、CPn分居呼吸道病毒感染的前三位;除CPn以秋季感染明显高于春冬夏季节;其余病毒感染无明显季节、年龄、性别差异.     

表达RSVF或G蛋白的单剂重组副流感病毒5型载体疫苗使棉鼠和非洲绿猴抵抗了RSV的攻击

正人呼吸道合胞病毒(RSV)是在婴幼儿、免疫功能低下者和老年人中引起严重呼吸系统疾病的常见病因。目前没有可用疫苗。在此研究中,作者用棉鼠和非洲绿猴模型评估了表达RSV F(PIV5/F)或G(PIV5/G)蛋白的两种副流感病毒5型(PIV5)-载体疫苗的复制、免疫原性和抗RSV攻击的保护功效。经过单次鼻内接种后,两种动物短期排泄了疫苗的病毒但没有明显的临床症状。     

Impact of non-pharmaceutical interventions during the COVID-19 pandemic on common childhood respiratory viruses – An epidemiological study based on hospital data

Considering common childhood respiratory viruses and SARS-CoV-2 share similar transmission routes, non-pharmaceutical interventions (NPIs) to prevent SARS-CoV-2 may affect the epidemiology of respiratory viruses. Therefore, our study aimed to observe the epidemiologic characteristics of common childhood respiratory viruses in 2020 (after the pandemic) compared with 2019 (before the pandemic) in Hangzhou, China. The data were compared between 2019 and 2020 based on age and month, respectively. One or more viruses were detected in 3135/21452 (14.61%) specimens in 2019, which was significantly lower in 1110/8202 (13.53%) specimens in 2020. Respiratory syncytial virus (RSV) was the most commonly detected virus in 2019 and 2020. The positive rate of adenovirus (ADV), parainfluenza virus (PIV)1, PIV2, and PIV3 in 2020 was significantly decreased in 2019. In 2020, RSV replaced ADV as the most predominant virus in children aged 1–6 years, and the positive rate of influenza virus A (FluA), influenza virus B (FluB), PIV1, and PIV2 was not correlated to age. FluA, FluB, and PIV2 were not almost detected from February 2020. The positive rates of ADV and PIV1 were uncorrelated to the month in 2020. By strict NPIs, besides controlling the COVID-19 pandemic, incredible progress has been made to reduce the prevalence of common childhood respiratory viruses.     

Plasma interferon-gamma, interleukin-10 and soluble markers of immune activation in infants with primary adenovirus (ADV) and respiratory syncytial virus (RSV) infection

Adenovirus (ADV) and respiratory syncytial virus (RSV) are etiological agents of acute respiratory tract infection in infants. Long-term prognosis of ADV infection includes severe lung damage, bronchiectasis and hyperlucent lung, while RSV infection is associated with development of recurrent wheezing and subsequent asthma. These differences may be related to differences in the primary immune responses elicited by these viruses. In this paper, we investigated the type of cytokine responses and the magnitude of immune activation in ADV and RSV infections in infants. We examined plasma concentrations of interferon-gamma (IFN-gamma), interleukin-10 (IL-10), soluble interleukin-2 receptor (sCD25) and soluble tumor necrosis factor receptor II (sTNFR-II) in previously healthy infants during the acute phase of primary ADV infection (n = 21) and RSV infection (n = 68), and in uninfected controls (n = 44). In ADV-infected infants, IFN-gamma plasma levels were significantly higher than those observed in RSV cases and the control group (p < 0.05). RSV cases did not show any differences in IFN-gamma plasma levels compared to the other groups. sCD25 levels were significantly higher in ADV- and RSV-infected infants than in controls (p < 0.0001), and higher in ADV than in RSV cases (p < 0.05). sTNFR-II levels were significantly higher in RSV- and ADV-infected infants than in controls (p < 0.0001, p < 0.05, respectively), and higher in RSV than in ADV infection (p < 0.05). No significant differences were observed in IL-10 plasma concentrations between the three groups. These results indicate that ADV and RSV infections in infants differ significantly with regard to the magnitude of production of interferon-gamma and soluble immune activation markers sCD25 and sTNFR-II. These immunological differences may be involved in the different clinical outcomes associated with these viral infections.     

五种主要上呼吸道病毒多重RT-PCR 检测方法的建立

目的:建立甲型、乙型流感病毒、呼吸道合胞病毒A型、B型(RSV-A、RSV-B)和腺病毒(ADV)五种主要上呼吸道病毒的多重RT-PCR检测方法。方法:利用Primer premier5.0分别针对甲型流感病毒的M基因、乙型流感病毒的PB1基因、RSV-A和RSV-B的F基因及ADV的hexon基因设计五对特异性引物,对Mg2+、dNTP、引物浓度及退火温度等进行优化,建立同时检测甲型、乙型流感病毒、RSV-A、RSV-B和ADV的多重RT-PCR方法,并验证该检测方法的灵敏性。结果:所建立的五种病毒的多重RT-PCR方法可以同时或者分别扩增甲型、乙型流感病毒、RSV-A、RSV-B及ADV的141bp、635bp、525bp、377b和283bp基因片段,敏感度分别达到770PFU/ml、800PFU/ml、680PFU/ml、970PFU/ml和850PFU/ml,且五种病毒间无交叉反应。结论:所建立的多重RT-PCR方法可以迅速准确地检测甲型、乙型流感病毒、RSV-A、RSV-B和ADV,为五种病毒的检测提供了一种方便易行的方法。     

Respiratory viruses in children younger than five years old with acute respiratory disease from 2001 to 2004 in Uberlândia, MG, Brazil

The main viruses involved in acute respiratory diseases among children are: respiratory syncytial virus (RSV), influenzavirus (FLU), parainfluenzavirus (PIV), adenovirus (AdV), human rhinovirus (HRV), and the human metapneumovirus (hMPV). The purpose of the present study was to identify respiratory viruses that affected children younger than five years old in Uberlandia, Midwestern Brazil. Nasopharyngeal aspirates from 379 children attended at Hospital de Clínicas (HC/UFU), from 2001 to 2004, with acute respiratory disease, were collected and tested by immunofluorescence assay (IFA) to detect RSV, FLU A and B, PIV 1, 2, and 3 and AdV, and RT-PCR to detect HRV. RSV was detected in 26.4% (100/379) of samples, FLU A and B in 9.5% (36/379), PIV 1, 2 and 3 in 6.3% (24/379) and AdV in 3.7% (14/379). HRV were detected in 29.6% (112/379) of the negative and indeterminate samples tested by IFI. RSV, particularly among children less than six months of life, and HRV cases showed highest incidence. Negative samples by both IFA and RT-PCR might reflect the presence of other pathogens, such as hMPV, coronavirus, and bacteria. Laboratorial diagnosis constituted an essential instrument to determine the incidence of the most common viruses in respiratory infections among children in this region.     

Quantitative proteome profiling of respiratory virus-infected lung epithelial cells

Respiratory virus infections are among the primary causes of morbidity and mortality in humans. Influenza virus, respiratory syncytial virus (RSV), parainfluenza (PIV) and human metapneumovirus (hMPV) are major causes of respiratory illness in humans. Especially young children and the elderly are susceptible to infections with these viruses. In this study we aim to gain detailed insight into the molecular pathogenesis of respiratory virus infections by studying the protein expression profiles of infected lung epithelial cells.A549 cells were exposed to a set of respiratory viruses [RSV, hMPV, PIV and Measles virus (MV)] using both live and UV-inactivated virus preparations. Cells were harvested at different time points after infection and processed for proteomics analysis by 2-dimensional difference gel electrophoresis. Samples derived from infected cells were compared to mock-infected cells to identify proteins that are differentially expressed due to infection.We show that RSV, hMPV, PIV3, and MV induced similar core host responses and that mainly proteins involved in defense against ER stress and apoptosis were affected which points towards an induction of apoptosis upon infection. By 2-D DIGE analyses we have gathered information on the induction of apoptosis by respiratory viruses in A549 cells.     

Influenza and Other Respiratory Viruses Detected by Influenza-Like Illness Surveillance in Leyte Island,the Philippines, 2010–2013

This study aimed to determine the role of influenza-like illness (ILI) surveillance conducted on Leyte Island, the Philippines, including involvement of other respiratory viruses, from 2010 to 2013. ILI surveillance was conducted from January 2010 to March 2013 with 3 sentinel sites located in Tacloban city, Palo and Tanauan of Leyte Island. ILI was defined as fever ≥38°C or feverish feeling and either cough or running nose in a patient of any age. Influenza virus and other 5 respiratory viruses were searched. A total of 5,550 ILI cases visited the 3 sites and specimens were collected from 2,031 (36.6%) cases. Among the cases sampled, 1,637 (75.6%) were children aged <5 years. 874 (43.0%) cases were positive for at least one of the respiratory viruses tested. Influenza virus and respiratory syncytial virus (RSV) were predominantly detected (both were 25.7%) followed by human rhinovirus (HRV) (17.5%). The age distributions were significantly different between those who were positive for influenza, HRV, and RSV. ILI cases were reported throughout the year and influenza virus was co-detected with those viruses on approximately half of the weeks of study period (RSV in 60.5% and HRV 47.4%). In terms of clinical manifestations, only the rates of headache and sore throat were significantly higher in influenza positive cases than cases positive to other viruses. In conclusion, syndromic ILI surveillance in this area is difficult to detect the start of influenza epidemic without laboratory confirmation which requires huge resources. Age was an important factor that affected positive rates of influenza and other respiratory viruses. Involvement of older age children may be useful to detect influenza more effectively.