西藏地区儿童急性呼吸道感染中呼吸道合胞病毒的初步研究

本研究为了解西藏地区儿童急性呼吸道感染中呼吸道合胞病毒(Respiratory syncytial virus,RSV)及基因型别。首先采用直接免疫荧光法检测2011年4～7月西藏自治区人民医院儿科病房因急性呼吸道感染住院患儿的鼻咽分泌物标本中7种常见的呼吸道病毒及人类偏肺病毒(Human metapneumovirus,hMPV)的抗原。然后对RSV抗原阳性的标本分别提取RNA,用逆转录-巢式聚合酶链反应法(Nest-PCR)确定RSV型别,同时用实时荧光PCR(Real-Time PCR)方法进行验证。再通过对G蛋白基因PCR扩增产物序列测定确定RSV的基因型。通过与GenBank中不同地区RSV分离株的G蛋白基因序列比对,了解西藏地区RSV G蛋白的结构特点及变异情况。结果表明,从167例标本中检测出呼吸道病毒抗原阳性的为65例,总阳性率为38.9%(65/167),其中RSV 45例,占阳性标本的69.2%(45/65),对其中42例RSV阳性标本进行了PCR分型,其中40例为A亚型,2例为B亚型。对7株A亚型RSV G蛋白基因PCR产物测序结果显示,全部为GA2基因型。西藏RSV与RSV原型株A2株核苷酸的同源性为90.7%～91.8%,氨基酸的同源性只有86.5%～87.2%。氨基酸的变异主要集中在胞外区一个高度保守序列的两端。7株西藏A亚型RSV G蛋白的核苷酸序列与GenBank中不同的RSV分离株相比同源性为90.7%～91.8%。西藏地区2011年春季小儿急性呼吸道感染的病毒病原主要为呼吸道合胞病毒,A亚型是2011年西藏地区的流行优势型别,其G蛋白胞外区基因具有较高的变异性。     

2017-2020年河南省漯河市呼吸道合胞病毒流行特征研究

呼吸道合胞病毒(Respiratory syncytial virus,RSV)是引起严重急性呼吸道感染(Severe acute respiratory infection,SARI)的一个重要病原,尤其以5岁以下儿童为主.为了解河南省漯河市SARI住院患者中RSV感染的流行病学和临床特征,为RSV预防控制及临床诊疗提供科学数据,本研究采集2017年10月至2020年8月河南省漯河市SARI住院病例的咽拭子,并收集流行病学和临床信息.采用荧光定量PCR方法鉴定RSV A/B阳性病例,分析其流行病学和临床特征.结果显示,本研究共入组1335例SARI病例,其中220例(16.48％)为RSV阳性,A和B亚型分别占64.55％和30.45％.RSV感染以5岁以下儿童为主(占91.36％),2岁以下婴幼儿占RSV感染病例的一半以上(占55.37％).RSV流行高峰出现在11月-次年1月,不同年份流行季可前后相差一个月.A﹑B亚型在不同月份可单独流行也可共流行.受新型冠状病毒肺炎疫情影响,2020年2-8月SARI病例数较往年同期减少60％以上,RSV阳性率在2020年2-8月降低为0.与非RSV感染组相比,RSV更易感染2岁以下儿童,下呼吸道感染占比更高(以支气管肺炎为主).本研究通过近3年SARI病例监测,揭示河南省漯河市RSV感染以冬春季常见,以下呼吸道感染为主,主要感染5岁以下儿童,其中2岁以下婴幼儿是防控重点人群.新型冠状病毒肺炎流行期间,由于限制性防控措施的干预,RSV感染大大降低.本研究将为RSV疫苗和单克隆抗体等预防性干预手段的使用策略提供基础数据.     

急性下呼吸道感染患儿中RSV的分离鉴定及分析

目的对2015年兰州单中心急性下呼吸道感染患儿中呼吸道合胞病毒(respiratory syncytial virus,RSV)进行分离鉴定及分析。方法收集临床急性下呼吸道感染患儿痰分泌物226份,在HEP-2细胞中连续盲传培养3代,收获的培养物用半巢式RT-PCR进行检测,选取强阳性样品进行测序以确定其型别,并对其流行病学特征进行分析。结果 226份样品中RSV阳性样本为76份(阳性率为34%)。RSV阳性患儿中,男女性别比为3∶1;年龄分布6月龄患儿占45%,6月龄~2岁占35%,2~5岁的占20%。冬季为发病高峰。76例RSV阳性患儿中,临床诊断依次为毛细支气管炎31例(40.79%)、喘息性支气管炎21例(27.36%)、支气管肺炎20例(26.32%)和支气管哮喘4例(5.26%)。40份强阳性RSV样品经测序确定为B亚型。结论 RSV是引起婴幼儿急性下呼吸道感染的重要病原之一,2015年RSV的流行株主要为B亚型。     

住院肺炎患儿偏肺病毒和呼吸道合胞病毒感染研究

为探讨沈阳地区肺炎患儿中偏肺病毒(hMPV)和呼吸道合胞病毒(RSV)感染情况,用反转录聚合酶链反应(RT-PCR)法对部分因肺部感染住院患儿进行了病原学研究。结果显示,hMPV感染率为9%;而RSV感染率为46%,其中A型占40%,B型占60%。研究结果表明,在婴幼儿肺炎患儿中RSV感染率高于hMPV感染率,两者所致肺炎在发病年龄、性别及临床症状上无显著区别。     

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。     

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。     

呼吸道合胞病毒疫苗研究进展

呼吸道合胞病毒(RSV)是引起严重下呼吸道感染的重要病原体,尽管经历了半个多世纪的努力,至今仍未有安全有效的RSV疫苗上市。近年来在RSV F蛋白结构生物学方面的研究进展为新一代RSV疫苗的开发提供了新方向,同时更多的采用不同技术、或针对不同人群的RSV侯选疫苗也在迅速发展,尤其是针对婴幼儿及老年人的RSV侯选苗已有60多种在研究中,大部分已处于临床前研究阶段,18种侯选苗已进入临床试验。我们简要介绍RSV疫苗的最新研究进展。     

呼吸道合胞病毒非结构蛋白NS1、NS2的研究进展

呼吸道合胞病毒(reespiratory syncytial virus,RSV)是引起婴幼儿和老年人下呼吸道感染的重要病原体之一.由于该病毒的致病机理还不太清楚导致目前尚无有效治疗RSV的方法.研究表明,呼吸道合胞病毒的非结构蛋白NS1、NS2具有抗细胞凋亡的作用,同时可以逃避宿主免疫系统(IFN)对病毒的干扰,有利于病毒复制.敲除这两种基因的减毒活疫苗和袁达沉默NS1的小干扰RNA(siRNA)的质粒研究已经取得了一定的进展.对非结构蛋白功能的深入研究有助于了解RSV的致病机理,同时为预防和治疗RSV感染奠定理论基础.     

136例急性呼吸道感染患儿病原菌特点及鼻咽分泌物中呼吸道合胞病毒和人偏肺病毒基因分析

目的分析136例急性呼吸道感染患儿的病原菌特点,并对患儿鼻咽分泌物中呼吸道合胞病毒(RSV)、人偏肺病毒(hMPV)进行检测,为后续研究提供参考。方法选择2017年1月至2019年4月我院收治的136例急性呼吸道感染患儿为研究对象,分析患儿呼吸道感染病原菌分布、耐药性,并对不同性别、年龄和感染部位患儿鼻咽分泌物中hMPV和RSV基因进行检测。结果136例急性呼吸道感染患儿共检出病原菌164株,其中革兰阴性菌占70.73%(116/164),革兰阳性菌占25.00%(41/164),真菌占4.27%(7/164);以铜绿假单胞菌(26.22%)、肺炎克雷伯菌(17.07%)、大肠埃希菌(15.24%)、金黄色葡萄球菌(12.20%)为主。药敏试验显示,铜绿假单胞菌、肺炎克雷伯菌、大肠埃希菌均对哌拉西林/他唑巴坦、头孢哌哃/舒巴坦、亚胺培南、左氧氟沙星、阿米卡星的耐药性较低(耐药率<50.0%)。金黄色葡萄球菌对万古霉素、呋喃妥因的耐药率为0.00%,对克林霉素、利福平的耐药率低于50.00%。136例患儿共检出hMPV 21例(15.44%),RSV 33例(24.26%)。不同性别患儿hMPV、RSV检出情况差异无统计学意义(均P>0.05)。1~2岁患儿hMPV检出率最高,0~5岁患儿RSV检出率较高。喘息性支气管炎、毛细支气管炎患儿hMPV检出率均较高,支气管肺炎、毛细支气管炎患儿RSV检出率均较高。结论急性呼吸道感染患儿病原菌以革兰阴性菌为主,应结合病原菌的药敏试验结果为患儿选择更有针对性的药物进行治疗。此外,应重视患儿RSV、hMPV感染情况,采取科学有效的方式对急性呼吸道感染进行预防和治疗。     

Association of Respiratory Viruses with Outcomes of Severe Childhood Pneumonia in Botswana

BackgroundThe highest incidence of childhood acute lower respiratory tract infection (ALRI) is in low- and middle-income countries. Few studies examined whether detection of respiratory viruses predicts ALRI outcomes in these settings.MethodsWe conducted prospective cohort and case-control studies of children 1-23 months of age in Botswana. Cases met clinical criteria for pneumonia and were recruited within six hours of presentation to a referral hospital. Controls were children without pneumonia matched to cases by primary care clinic and date of enrollment. Nasopharyngeal specimens were tested for respiratory viruses using polymerase chain reaction. We compared detection rates of specific viruses in matched case-control pairs. We examined the effect of respiratory syncytial virus (RSV) and other respiratory viruses on pneumonia outcomes.ResultsBetween April 2012 and August 2014, we enrolled 310 cases, of which 133 had matched controls. Median ages of cases and controls were 6.1 and 6.4 months, respectively. One or more viruses were detected from 75% of cases and 34% of controls. RSV and human metapneumovirus were more frequent among cases than controls, but only enterovirus/rhinovirus was detected from asymptomatic controls. Compared with non-RSV viruses, RSV was associated with an increased risk of treatment failure at 48 hours [risk ratio (RR): 1.85; 95% confidence interval (CI): 1.20, 2.84], more days of respiratory support [mean difference (MD): 1.26 days; 95% CI: 0.30, 2.22 days], and longer duration of hospitalization [MD: 1.35 days; 95% CI: 0.20, 2.50 days], but lower in-hospital mortality [RR: 0.09; 95% CI: 0.01, 0.80] in children with pneumonia.ConclusionsRespiratory viruses were detected from most children hospitalized with ALRI in Botswana, but only RSV and human metapneumovirus were more frequent than among children without ALRI. Detection of RSV from children with ALRI predicted a protracted illness course but lower mortality compared with non-RSV viruses.     

单克隆抗体在呼吸道合胞病毒感染诊断和防治中的应用

呼吸道合胞病毒 (RSV)是世界范围内婴幼儿下呼吸道感染的重要病原 ,目前尚无成熟疫苗应用于临床。展开对RSV被动免疫的研究显得尤为重要。单克隆抗体成为深入研究和防治RSV感染的有力武器。本文综述了单克隆抗体在RSV感染的诊断、预防和治疗中的应用     

Clinical and Epidemiologic Characteristics of Hospitalized Patients with Laboratory-Confirmed Respiratory Syncytial Virus Infection in Eastern China between 2009 and 2013: A Retrospective Study

Respiratory syncytial virus (RSV) is a leading cause of morbidity and mortality worldwide in children aged <5 years and older adults with acute lower respiratory infections (ALRIs). However, few studies regarding the epidemiology of hospitalizations for RSV infection have been performed previously in China. Here, we aimed to describe the clinical and epidemiologic characteristics of hospitalized patients with laboratory-confirmed RSV infection in eastern China. Active surveillance for hospitalized ALRI patients using a broad case definition based on symptoms was performed from 2009–2013 in 12 sentinel hospitals in eastern China. Clinical and epidemiologic data pertaining to hospitalized patients of all ages with laboratory-confirmed RSV infection by PCR assay were collected and analyzed in this study. From 2009 to 2013, 1046 hospitalized patients with laboratory-confirmed RSV infection were enrolled in this study, and 14.7% of patients had subtype A, 24.2% of patients had subtype B, 23.8% of patients with subtype not performed, and 37.3% of patients had RSV coinfections with other viruses. RSV and influenza coinfections (33.3%) were the most common coinfections noted in this study. Moreover, young children aged <5 years (89.1%, 932/1046), particularly young infants aged <1 year (43.3%, 453/1046), represented the highest proportion of patients with RSV infections. In contrast, older adults aged ≥60 years (1.1%, 12/1046) represented the lowest proportion of patients with RSV infections among enrolled patients. The peak RSV infection period occurred mainly during autumn and winter, and 57% and 66% of patients exhibited symptoms such as fever (body temperature ≥38°C) and cough separately. Additionally, only a small number of patients were treated with broad-spectrum antiviral drugs, and most of patients were treated with antimicrobial drugs that were not appropriate for RSV infection. RSV is a leading viral pathogen and a common cause of viral infection in young children aged <5 years with ALRIs in eastern China. Effective vaccines and antiviral agents targeting RSV are needed to mitigate its large public health impact.     

A rhesus monkey model of respiratory syncytial virus infection

Respiratory syncytial virus (RSV) is a major cause of lower respiratory tract disease in infants and young children worldwide. To date, there is no single animal model that adequately reproduces all human disease states. Here, we have developed a model of experimental infection with human RSV in infant Rhesus macaques. Infected animals demonstrated mild clinical disease including increased respiratory rates, fever and adventitious lung sounds. While more severe disease was not observed, preliminary virological and histopathological findings are promising. It is anticipated that with further optimization, this model will provide a useful system with which to study disease due to RSV infection and evaluate candidate vaccines.     

A multi-center study on Molecular Epidemiology of Human Respiratory Syncytial Virus from Children with Acute Lower Respiratory Tract Infections in the Mainland of China between 2015 and 2019

Virologica Sinica - Human respiratory syncytial virus (RSV) is a major pathogen of acute lower respiratory tract infection among young children. To investigate the prevalence and genetic...     

Prevalence of Herpes and Respiratory Viruses in Induced Sputum among Hospitalized Children with Non Typical Bacterial Community-Acquired Pneumonia

Objective

Few comprehensive studies have searched for viruses in infants and young children with community-acquired pneumonia (CAP) in China. The aim of this study was to investigate the roles of human herpes viruses (HHVs) and other respiratory viruses in CAP not caused by typical bacterial infection and to determine their prevalence and clinical significance.

Methods

Induced sputum (IS) samples were collected from 354 hospitalised patients (infants, n = 205; children, n = 149) with respiratory illness (CAP or non-CAP) admitted to Wenling Hospital of China. We tested for HHVs and respiratory viruses using PCR-based assays. The epidemiological profiles were also analysed.

Results

High rate of virus detection (more than 98%) and co-infection (more than 80%) were found among IS samples from 354 hospitalised infants and children with respiratory illness in this study. Of 273 CAP samples tested, CMV (91.6%), HHV-6 (50.9%), RSV (37.4%), EBV (35.5%), HBoV (28.2%), HHV-7 (18.3%) and rhinovirus (17.2%) were the most commonly detected viruses. Of 81 non- CAP samples tested, CMV (63%), RSV (49.4%), HHV-6 (42%), EBV (24.7%), HHV-7 (13.6%) and HBoV (8.6%) were the dominant viruses detected. The prevalence of several viral agents (rhinovirus, bocavirus, adenovirus and CMV) among IS samples of CAP were significantly higher than that of non-CAP control group. We also found the prevalence of RSV coinfection with HHVs was also higher among CAP group than that of non-CAP control.

Conclusions

With sensitive molecular detection techniques and IS samples, high rates of viral identification were achieved in infants and young children with respiratory illness in a rural area of China. The clinical significance of rhinovirus, bocavirus, adenovirus and HHV (especially CMV) infections should receive greater attention in future treatment and prevention studies of CAP in infants and children.     

Infections caused by RSV among children and adults during two epidemic seasons

Respiratory Syncytial Virus (RSV) is one of the most common causes of lower respiratory tract infections in young children, immunocompromised patients (children and adults), patients with chronic respiratory diseases and elderly people. Reinfections occur throughout the life, but the severity of disease decreased with subsequent infection. The aim of this study was to analyze the frequency of RSV infections in two selected subpopulations: young children (below 5 y.) and adults with chronic respiratory diseases (25-87 y.). Nasopharyngeal swabs (334) collected from October 2008 to March 2010 were examined. The presence of RSV genome was determined by RT-PCR and the presence of RSV antigen by quick immunochromatographic test. Positive results of RT-PCR were found in 45.2% of all swabs: 48.6% samples in 2008; 41.5% in 2009; 50.8% in 2010. The highest frequency of RSV-positive samples was in fall-winter months, but differences in RSV epidemic seasons were found. In the first season (2008-2009) an increased number of RSV infections was observed from November 2008, but in the second season--from January 2010. Generally, the frequency of RSV-positive RT-PCR among children was 53%, among adults 25%. The highest difference was observed in the first three-month period of 2010. RT-PCR positive samples were found in 68.5% of children and 5.9% of adults. However, the RSV antigen was found in 44.4% of samples collected from adults in this period. Our results indicate that the contribution of RSV infections during epidemic season of respiratory tract infections in Poland was really high among children and adults.     

Association of TLR4 polymorphisms with symptomatic respiratory syncytial virus infection in high-risk infants and young children

Respiratory syncytial virus (RSV) is a leading cause of infant mortality worldwide. Although anti-RSV Ab prophylaxis has greatly reduced infant mortality in the United States, there is currently no vaccine or effective antiviral therapy. RSV fusion (F) protein activates cells through TLR4. Two single nucleotide polymorphisms (SNPs) encoding Asp299Gly and Thr399Ile substitutions in the TLR4 ectodomain were previously associated with TLR4 hyporesponsiveness and increased susceptibility to bacterial infection. Prevalence of these SNPs was analyzed in a case series of 105 DNA samples extracted from archived nasal lavage samples from high-risk infants/young children with confirmed RSV disease who participated in two seminal clinical trials for anti-RSV prophylaxis. Frequencies of TLR4 SNPs in the case series were compared with those of literature controls, healthy adults, infants, and young children who presented with symptoms of respiratory infections (but not preselected for high risk for RSV). Both SNPs were highly associated with symptomatic RSV disease in this largely premature population (p < 0.0001), with 89.5% and 87.6% of cases being heterozygous for Asp299Gly and Thr399Ile polymorphisms versus published control frequencies of 10.5% and 6.5%, respectively. The other two control groups had similarly low frequencies. Our data suggest that heterozygosity of these two extracellular TLR4 polymorphisms is highly associated with symptomatic RSV disease in high-risk infants and support a dual role for TLR4 SNPs in prematurity and increased susceptibility to RSV not revealed by analysis of either alone.     

人呼吸道合胞病毒活疫苗研究进展

人呼吸道合胞病毒是引起婴幼儿支气管炎和肺炎的主要原因,也可导致免疫缺陷病人及老年人群显著发病和死亡.人呼吸道合胞病毒疫苗已被世界卫生组织(World Health Organization,WHO)列为全球最优先发展的疫苗之一.经过50多年的研究,尤其是随着重组技术和反向遗传学的出现,对RSV疫苗的研究取得了重要进展,...