6岁以下儿童急性下呼吸道感染的病毒病原及临床研究

目的　了解北京地区 6岁以下儿童中急性下呼吸道感染 (ALRI)患儿的病毒感染状况 ;探讨人类偏肺病毒 (hMPV)感染在儿童ALRI中的作用及所致支气管肺炎的临床特征。方法　 (1)采集 2 0 0 1年 2月至 2 0 0 3年 3月就诊于首都儿科研究所 4 4 6例年龄 1个月至 5岁的ALRI住院患儿的鼻咽分泌物 ,同时应用间接免疫荧光 (IFA)及病毒分离方法进行人类呼吸道合胞病毒 (hRSV)、流感病毒 (甲、乙型 )、副流感病毒 (Ⅰ、Ⅱ、Ⅲ型 )及腺病毒等 7种常见呼吸道病毒的病原检测。 (2 )采用逆转录聚合酶链反应 (RT PCR)方法对 2 0 0 2年 11月至 2 0 0 3年 3月住我院内科的ALRI患儿中经 7种呼吸道常见病毒病原检测阴性的 2 16份鼻咽洗液 ,进行了hMPV基因检测 ,并分析hMPV感染患儿的临床资料。结果　 (1) 4 4 6例患儿中 7种常见呼吸道病毒总阳性例数为 2 0 9例 (46 86 % ) ,hRSV阳性者最多为 16 3例 (36 5 5 % )。 (2 )在 2 16份上述 7种呼吸道病原检测阴性的标本中 6 9份测到hMPV阳性扩增产物 (31 9% ) ,占同期送检标本的 19 17%。结论　 (1)hRSV是北京地区冬春季节婴幼儿ALRI的主要病原。 (2 )2 0 0 2年 11月至 2 0 0 3年 3月我院ALRI住院患儿中有 19 17%与hMPV感染有关     

重庆地区急性下呼吸道感染住院患儿人偏肺病毒感染的流行病学研究

目的了解重庆地区急性下呼吸道感染(ALRTI)住院患儿人偏肺病毒(hMPV)感染的流行病学特点和临床特征。方法收集2006年4月至2008年3月重庆医科大学附属儿童医院呼吸科病房每周一、三、五因ALRTI住院患儿住院当日白天的鼻咽吸取物(NPA),采用荧光定量PCR方法检测hMPV基因组RNA,阳性标本采用传统PCR方法扩增F基因,进一步确定是否存在hMPV感染,扩增G基因用于遗传进化分析。同时采用传统PCR方法检测呼吸道合胞病毒(RSV)和冠状病毒NL-63(HCoV-NL63),直接免疫荧光法(DFA)检测流感病毒A、B亚型,副流感病毒1、2、3型和腺病毒。分析hMPV感染的流行病学特点和临床特征。结果研究期间共收集878份标本,占同期ALRTI入院病例的13.9%(878/6296例)。hMPV阳性227/878例(25.9%)。男∶女为1.9∶1。hMPV阳性227例中,6个月119例(52.4%),～2岁72例(31.7%),～6岁34例(15.0%),6岁2例(0.9%)。hMPV流行特点为全年散发,好发季节为冬春季。hMPV阳性率2006年4月至2007年3月为18.5%(72/390例),2007年4月至2008年3月为31.8%(155/488例)。227例hMPV阳性患儿临床诊断以肺炎(148例,65.2%)和毛细支气管炎(65例,28.6%)为主。hMPV可与常见呼吸道病毒协同感染,其中与RSV协同感染率最高(41.9%,95/227例)。遗传进化分析显示,研究期间重庆地区hMPVA和B基因型均流行,以A2亚型为优势株。结论 hMPV是重庆地区ALRTI住院患儿重要的呼吸道病毒病原之一,以6个月婴儿为主要感染人群。     

重庆地区人类偏肺病毒感染与小儿毛细支气管炎

目的探讨人类偏肺病毒(human metapneumovirus，hMPV)在重庆地区小儿毛细支气管炎中的感染情况，比较hMPV与呼吸道合胞病毒(RSV)感染引起的毛细支气管炎的临床特点。方法用免疫荧光快速诊断方法，检测80例毛细支气管炎患儿鼻咽分泌物中RSV、腺病毒(Ado)、流感病毒(Flu)A、B、副流感病毒(Para)1、2、3七种常见呼吸道病毒，采用RT-PCR的方法检测上述患儿鼻咽分泌物中hMPV的L基因和M基因。结果80例毛细支气管炎患儿鼻咽分泌物中病毒检出率为65％，在病毒阳性检出标本中RSV感染占63．4％(33／52例)，Flu-B(3／52例)和Para-3(3／52例)均占5．77％，Para-1占1．92％(1／52例)，Flu-A与Flu-B混合感染占3．85％(2／52例)；80例患儿的鼻咽分泌物中发现10例为hMPVL和M基因同时阳性，其中单独hMPV阳性7例，3例为RSV与hMPV合并感染，hMPV阳性占病毒阳性检出标本的13．46％(7／52例)，hMPV与RSV合并感染为5．77％(3／52例)。结论RSV仍然是引起毛细支气管炎的首位病原，约为41．25％；hMPV感染在重庆地区小儿毛细支气管炎中居第二位，约占8．75％(7／80例)，仅从临床表现上很难鉴别hMPV感染与RSV感染。     

2006-2007年苏州地区儿童呼吸道人类偏肺病毒感染的流行和临床特征

目的 了解苏州地区儿童人类偏肺病毒(hMPV)呼吸道感染的流行情况及临床特征.方法 收集2006年1月-2007年12月我院呼吸科4702例急性呼吸道感染住院患儿的呼吸道分泌物,用逆转录-聚合酶链反应法检测hMPV N基因,随机挑选部分阳性PCR扩增产物进行核苷酸测定并将所测序列在GenBank中进行比较分析.结果 4702例标本中,检测到376份RT-PCR阳性扩增产物,阳性率为8%;2006,2007年hMPV检测阳性率分别为8.4%、7.6%.hMPV检测阳性率分别在1、2、3月和11、12月呈现高峰,hMPV感染患儿的平均年龄为22.56个月,376例hMPV感染患儿中临床诊断为上呼吸道感染12例(3.2%);喉炎8例(2.1%);毛细支气管炎102例(27.1%);肺炎210例(55.9%);哮喘急性发作44例(11.7%).20份hMPV N基因部分核苷酸片断与GenBank中hMPVN基因序列同源性为99%～100%.结论 (1)苏州地区部分儿童的呼吸道感染与hMPV有关.(2)全年均存在hMPV感染,流行高峰在冬春季.(3)hMPV感染的临床特征无特异性.     

重庆地区急性呼吸道感染患儿鼻咽吸取物标本中人偏肺病毒的分离研究

目的分离新近发现的儿童呼吸道病毒病原——人偏肺病毒（hMPV）,并分析其感染的临床表现。方法选取重庆医科大学儿童医院2004年12月至2005年7月因呼吸道感染住院的患儿,无菌采集鼻咽吸取物。离心后细胞沉淀用直接免疫荧光法检测呼吸道合胞病毒、腺病毒、流感病毒A／B、副流感病毒1、2、3型等常见呼吸道病毒病原。上清接种于Veto-E6和LLC-MK2细胞,进行hMPV分离,出现细胞病变者,扩增hMPV N、F基因,F基因扩增产物序列递交GenBank。收集并分析hMPV感染患儿的临床资料。结果收集的86份鼻咽吸取物中,有6份接种Veto-E6和LLC-MK2细胞后出现细胞病变,且用RT-PCR成功扩增出hMPV N、F基因片段,证实hMPV分离成功。在6例阳性标本中,2例收集于冬季,2例春季,2例夏季。6位患儿年龄都小于2岁,诊断为支气管肺炎（2／6）、毛细支气管炎（2／6）、婴幼儿哮喘（1／6）和上呼吸道感染（1／6）。副流感病毒3和腺病毒可为协同感染病毒病原。结论首次在中国内地成功分离获得6株hMPV,从活病毒水平证实hMPV是导致中国儿童下呼吸道感染的病毒病原,hMPV感染的临床表现与呼吸道合胞病毒感染相似。采用病毒分离技术,hMPV在呼吸道感染患儿中的检出率约为7％（6／86）。     

儿童呼吸道人类偏肺病毒呼吸道合胞病毒和流感病毒感染的临床特征比较

目的 了解人类偏肺病毒(hMPV)在苏州地区儿童呼吸道感粢中的感染情况;比较hMPV、呼吸道合胞病毒(RSV)和流感病毒(Inf)引起呼吸道感染的临床特征.方法 选取2005年11月至2006年10月苏州大学附属儿童医院呼吸科1932例急性呼吸道感染住院患儿.采用无茵负压吸引法采集新鲜痰液.直接免疫荧光法检测RSV,腺病毒,Inf A、B型,副流感病毒1、2、3型等7种常见呼吸道病毒;同时用逆转录.聚合酶链反应法(RTPCR)检测hMPV N基因,随机挑选部分阳性PCR扩增产物进行核苷酸测定并将所测序列进行比较分析.结果 1932例标本中hMPV阳性例数为128例(6.6%),其中hMPV合并其他呼吸道病毒感染5例(3.9%);hMPV检测阳性率在冬季呈现高峰;hMPV感染患儿的平均年龄为22.17月龄,较RSV和Inf感染患儿大;hMPV感染者临床特征与RSV和Inf感染相似,喘息在RSV和hMPV感染患儿中更为多见,发热则更多见于hMPV和Inf感染患儿.RSV感染后所致的首要疾病是毛细支气管炎,hMPV和Inf感染后所致的首要疾病是肺炎.单纯hMPV与hMPV合并其他呼吸道病毒感染临床表现差异无统计学意义(P＞0.05).结论 苏州地区部分儿童的呼吸道感染与hMPV有关;hMPV感染临床特征无特异性;hMPV在合并其他呼吸道病毒感染时并不加重病情.     

2006—2008年苏州地区儿童呼吸道人偏肺病毒感染的流行和临床特征

目的了解人类偏肺病毒(hMPV)在苏州地区小儿呼吸系统感染中的感染情况,分析其流行病学特点;探讨hMPV感染后的临床特征。方法选取2006年1月—2008年12月苏州大学附属儿童医院6 395例急性呼吸道感染住院患儿,总结临床特征;用逆转录-聚合酶链反应法(RT-PCR)检测痰液中hMPV N基因,随机挑选部分阳性PCR扩增产物进行核苷酸测定,并将所测序列在GenBank中进行比较分析。结果 6 395例标本中,检测到589份RT-PCR阳性扩增产物,检出率为9.2%,其中2006年度hMPV检出率为8.4%,2007年为7.6%,2008年为12.6%。hMPV在3年的每个月份都有检出,但在2006年12月、2007年1月和2008年5月呈现年度高峰,分别为24.2%、21.6%、31.9%。589例hMPV感染患儿的平均年龄为22.21个月;临床诊断上呼吸道感染3.6%,喉炎2.0%,毛细支气管炎20.2%,肺炎61.8%,哮喘急性发作12.4%。主要临床表现为咳嗽,鼻塞流涕,发热,呼吸困难,哺喂困难,喘息。20份hMPV N基因部分核苷酸片断与GenBank中hMPV N基因序列同源性为99%～100%。结论苏州地区部分儿童的呼吸道感染与hMPV有关。全年均存在hMPV感染,流行高峰在冬春季,连续3年的检出率有较大差异。hMPV感染的临床特征无特异性。     

急性呼吸道感染住院儿童人偏肺病毒感染的流行情况及临床特征

目的了解急性呼吸道感染住院患儿人偏肺病毒(hMPV)呼吸道感染的流行情况及临床特征。方法收集2006年11月～2007年2月本院儿科中心的112例急性呼吸道感染住院患儿的鼻咽分泌物标本,对其中65例用直接免疫荧光法(DFA)检测常见7种呼吸道病毒特异性抗原阴性患儿鼻咽分泌物用间接免疫荧光法(IFA)检测hMPV抗原。结果1.IFA能检测到鼻咽分泌物中hMPV抗原;2.阳性8例,阳性率为12.31%,112例中单一hMPV感染率为7.14%;3.hMPV的检出高峰主要在11月份及次年1月份;4.在≤5岁年龄组阳性率为15%(6/40例),>5岁年龄组阳性率为8%(2/25例),二组hMPV感染阳性率差异无统计学意义(P>0.05);5.hMPV感染的临床表现无特征性。结论西安地区儿童急性呼吸道感染与hMPV感染有关;咳嗽、发热是hMPV感染后的主要临床症状。     

急性下呼吸道感染患儿病毒病原学及其临床特征

目的 了解急件下呼吸道感染(ALRI)住院患儿病毒感染情况及其临床特征.方法 收集2004-2006年冬季(11月至次年2月)因ALRI在北京儿童医院住院治疗的＜5岁患儿鼻咽吸出物333份.男230例,女103例;年龄1个月～4.8岁.采用PCR技术和病毒分离方法对鼻咽吸出物进行呼吸道合胞病毒(RSV)、腺病毒(AdV)、副流感病毒(PIV)1～4型、鼻病毒(RhV)、人偏肺病毒(hMPV)、冠状病毒(HCoV)及人博卡病毒(HBoV)检测.采用人喉上皮癌细胞和非洲猴肾细胞对鼻咽吸出物进行病毒分离,对阳性结果进行序列测定和鉴定.结果 ALRI 333例患儿中,病毒检测阳性123份(36.94%),其中RSV阳性102例,AdV、HBoV阳性各16例,RhV阳性1例,其中12例为双重阳性.未检测到PIV、hMPV及HCoV.333份鼻回吸出物中病毒分离得剑11株病毒株.经PCR扩增及序列测定证实均为AdV,包括AdV1型2株,AdV2型、AdV3型、AdV7型各3株.肺炎患儿病毒阳性检出率为30.35%,毛细支气管炎患儿病毒阳性检出率为62.50%,急性支气管炎患儿病毒阳性检出率为23.08%,毛细支气管炎患儿病毒阳性检出率高于肺炎和急性支气管炎患儿(χ2=24.75,19.66 Pa＜0.01).结论 RSV感染在北京儿章医院2004-2006年冬季ALRI住院患儿中占首位;感染的AdV血清型包括AdV 1型、AdV 2型、AdV 3型及AdV 7型;HBoV可能是ALRI的重要病原.毛细支气管炎患儿病毒感染检出率最高.     

人类偏肺病毒毛细支气管炎患儿细胞免疫分析

目的 探讨人类偏肺病毒(human metapneumovirus,hMPV)感染致毛细支气管炎患儿细胞免疫功能的变化.方法 用RT-PCR方法对255例毛细支气管炎住院患儿的呼吸道分泌物进行hMPV N基因检测,以明确hMPV感染.采用流式细胞仪检测患儿外周血T淋巴细胞亚群B淋巴细胞和NK细胞,以30例健康儿童作为对照组.结果 共检测到51份RT-PCR阳性扩增产物,阳性率为20.0%;hMPV感染组外周血 CD3+T细胞、CD4+T细胞、CD8+T细胞与对照组比较差异无统计学意义,CD19+、CD19+CD23+、CD4+CD25+T细胞均明显增高(P<0.01),而NK细胞下降(P<0.05).结论 hMPV是毛细支气管炎的重要病原之一,hMPV感染后机体细胞免疫功能明显紊乱.     

Clinical course of hospitalised children infected with human metapneumovirus and respiratory syncytial virus

AIM: To describe the clinical presentation and outcomes of hospitalised patients infected with human metapneumovirus (hMPV) and human respiratory syncytial virus (hRSV) in a tertiary hospital in Cape Town, South Africa. METHODS: hMPV was identified in 17 respiratory specimens submitted for viral studies during the period 2001-2003. These patients' medical folders were retrospectively reviewed for clinical, radiological and laboratory data, together with a convenience sample of 20 hRSV-infected patients. RESULTS: hMPV-infected patients were older than those infected with hRSV (P = 0.04) and required a longer hospital stay (P = 0.02). Presenting clinical signs and symptoms were similar between groups. Fourteen (87.5%) hMPV- and 16 (80%) hRSV-infected patients presented with co-morbid and/or immunosuppressive conditions (P > or = 0.5). The most common abnormalities on chest radiographs in both groups were bronchial wall thickening, focal consolidation and atelectasis. Six (37.5%) hMPV- and 11 (55%) hRSV-infected patients required admission to the paediatric intensive care unit (P > 0.1) with five (31.3%) hMPV- and eight (40%) hRSV-infected patients requiring intubation and ventilation (P > 0.5). Three (18.7%) hMPV-patients and three (15%) hRSV-infected patients died during this admission (P > 0.5). All hMPV-infected patients who died had significant co-morbid conditions. CONCLUSIONS: These data confirm that hMPV is a significant respiratory pathogen in this setting, with similar presentation and outcome to hRSV infection. This is the largest report of hMPV infection causing significant morbidity, prolonged hospital stay and death, associated with underlying risk factors.     

Detection of new respiratory viruses in hospitalized infants with bronchiolitis: a three‐year prospective study

Aim: We have designed a study with the objective of describing the clinical impact of other viruses different from the respiratory syncytial virus (RSV) in hospitalized infants with bronchiolitis. Methods: A 3 year prospective study was conducted on infants admitted to the Paediatrics Department of the Severo Ochoa Hospital (Spain). We studied the frequency of 16 respiratory viruses. Clinical characteristics of RSV‐only infections were compared with other single agent viral infections. Results: Positive results were confirmed in 275 (86.5%) of the 318 children studied. A single virus was detected in 196 patients and 79 were dual or multiple viral infections. RSV was detected in 61.3% of total bronchiolitis. Rhinovirus (RV) was 17.4% of the identified virus, followed by human bocavirus (HBoV), adenovirus and metapneumovirus (hMPV). Only RV, HBoV and hMPV were significant as single infections. RSV patients were younger than HBoV (p > 0.0001) and hMPV (p = 0.025). Seasonality was clearly different between them. Children with RSV infection needed treatment in the intensive care unit more frequently than others. Conclusions: In hospitalized infants, RSV was the most frequent agent in bronchiolitis in winter, but other viruses were present in 47% of the patients. RV, HBoV and hMPV had a significant proportion of single infections. Clinical characteristics were similar amongst them, but seasonality was clearly different.     

太原地区急性呼吸道人偏肺病毒和人博卡病毒感染患儿临床特征分析

目的了解太原地区急性呼吸道感染(ARTIs)儿童人偏肺病毒(hMPV)与人博卡病毒(HBoV)的感染情况及其临床和流行病学特征。方法采集2012年11月—2013年5月及2013年11月—2014年5月就诊的ARTIs患儿549例,采集咽拭子标本,应用实时PCR方法检测hMPV与HBoV。结果 549例患儿的咽拭子标本中hMPV阳性56例(10.2%),HBoV阳性15例(2.7%)。其中2012年11月—2013年5月hMPV与HBoV检出率分别为12.3%和2.0%,2013年11月—2014年5月hMPV与HBoV检出率分别为6.5%和4.0%,两时间段hMPV检出率差异有统计学意义(P0.05),HBoV检出率差异无统计学意义(P0.05);不同月份hMPV、HBoV检出率差异无统计学意义(P0.05)。hMPV与HBoV均在2岁组中检出率最高。hMPV在喘息性支气管炎与毛细支气管炎患儿中检出率最高。结论太原地区hMPV和HBoV与部分儿童尤其是婴幼儿ARTIs有关,hMPV是诱发部分婴幼儿喘息性疾病的重要病原体之一。     

Outbreak of human metapneumovirus infection in norwegian children

INTRODUCTION: Human metapneumovirus (hMPV) was recently discovered in children with acute respiratory tract infection. We have studied the occurrence of hMPV and report clinical findings of 50 hMPV-infected children who were hospitalized during an outbreak in Norway. METHODS AND POPULATION: During 5 months from November 15, 2002 to April 14, 2003 we collected nasopharyngeal aspirate specimens from 236 children admitted because of respiratory tract infection (RTI). Samples were analyzed for influenza virus A/B, parainfluenza viruses 1, 2 and 3 and respiratory syncytial virus by direct immunofluorescence assays and cell culture. Rhinovirus, adenovirus and hMPV were identified by polymerase chain reaction. RESULTS: Human metapneumovirus was identified in 50 of 236 children (21%). Most (41 of 50) hMPV-infected children were hospitalized between November 15 and January 15, and during these 2 months hMPV was the most common isolate (41 of 72 isolates; 57%). Respiratory syncytial virus was identified in 36 children (15%), among whom 34 were admitted after the hMPV outbreak.The median age of hMPV-infected children was 12 months (range, 1 to 115 months), and one-half of the children had an underlying chronic disease. The most common symptoms were fever (86%), cough (90%), dyspnea (80%), wheezing (56%), rhinorrhea (44%), anorexia (48%) and vomiting (36%). Eight (16%) had an upper respiratory tract infection (rhinopharyngitis, n = 6; laryngitis, n = 2), 24 (48%) had bronchiolitis and 17 (34%) had pneumonia. Two-thirds with a lower RTI also had signs of upper RTI. Fourteen (28%) children needed supplemental oxygen, 1 was treated with continuous positive airway pressure and 2 were ventilated mechanically. CONCLUSION: Human metapneumovirus was the most common virus isolate during the winter season 2002 to 2003 in children hospitalized for respiratory tract infection. Upper respiratory tract infections and mild to severe bronchiolitis were most common, but a relatively high proportion of hospitalized children developed severe pneumonia.     

Human metapneumovirus infections cause similar symptoms and clinical severity as respiratory syncytial virus infections

Human metapneumovirus (hMPV) is a recently discovered pathogen in respiratory tract infection. The published literature suggests milder illness severity in hMPV compared with respiratory syncytial virus (RSV) infection. In two consecutive seasons, 637 nasopharyngeal aspirates from pediatric patients were tested by hMPV polymerase chain reaction, and risk factors and clinical and laboratory items were analyzed. The hMPV patients were compared with hMPV-negative but RSV-positive patients by matched pair analysis. HMPV was detected in 17.9% of all samples. In total, 88 hMPV-infected patients with complete datasets were considered. More than half of all hMPV patients were older than 12 months, 45.5% had at least one risk factor for a severe course of viral respiratory tract infection, and 27.3% were born prematurely, 15.9% with a birth weight <1,500 g. At least one other virus was also detected in 39 patients (44.3%; RSV in 29.5%). Coinfection did not result in greater severity of illness. On matched pair analysis (hMPV-positive/RSV-negative vs. hMPV-negative/RSV-positive), the epidemiological and clinical features of hMPV infection were similar to those of RSV infection, as in the hMPV group higher proportions of patients with hypoxemia on admission (33% vs. 21%) and of intensive care treatment (20.8% vs. 10.4%) were observed. More hMPV patients showed lobar infiltrates in radiological chest examination. In 60% of all hMPV infections, the attending physicians prescribed antimicrobial chemotherapy. We conclude that in hospitalized children, hMPV infection is as serious as RSV infection and therefore deserves the same attention. Virologic diagnosis from respiratory secretions is mandatory because clinical, laboratory, and radiological signs cannot sufficiently discriminate between viral and bacterial respiratory tract infection in infants and children.The first two authors contributed equally to this work.