呼吸道合胞病毒急性下呼吸道感染患儿的细胞免疫变化

呼吸道合胞病毒急性下呼吸道感染患儿的细胞免疫变化陈宗波，董永绥，赖丽霖呼吸道合胞病毒（ＲＳＶ）是引起小儿急性下呼吸道感染（ＡＬＲＩ）的重要病原。有关小儿ＲＳＶＡＬＲＩ时细胞免疫变化的多项指标综合报道国内不多，亦无对Ａ、Ｂ亚型ＲＳＶ感染进行观察，我们对...     

应用实时聚合酶链反应技术检测儿童呼吸道标本中的呼吸道合胞病毒

目的建立实时聚合酶链式反应（PCR）技术检测儿科呼吸道标本中呼吸道合胞病毒（RSV）的方法.方法（1）根据RSV N基因序列设计合成分别用于扩增RSV A亚型和B亚型的TaqMan探针和引物,建立实时PCR方法,进行灵敏度和特异性检测.（2）采用实时PCR检测61例RSV检测阳性的冻存标本和103例新收集的呼吸道标本,并与病毒分离、间接免疫荧光、巢式PCR结果相比较.结果（1）实时PCR对A亚型RSV检测的灵敏度为5.25 pfu,B亚型为3.75 pfu,与巢式PCR相同.（2）实时PCR检测其他呼吸道病毒阳性标本,结果均为阴性;A亚型和B亚型之间无交叉.（3）实时PCR检测其他方法检测过的RSV阳性的冻存标本61例,A亚型（30例）阳性为27例（90%）,B亚型（31）阳性为27例（87.1%）.（4）103例新鲜标本中,实时PCR检出RSV 35例（A亚型7,B亚型28）,阳性率34.0%,其中巢式PCR阳性31例,阳性率30.1%;间接免疫荧光阳性22例,阳性率21.4%;病毒分离阳性9例,阳性率8.7%.实时PCR阴性的68例,其他方法均为阴性.实时PCR与三种方法的一致性检验,一致率在74.76%～96.12%（P均＜0.01）;差异性检验结果说明,实时PCR阳性检出高于间接免疫荧光和病毒分离（P均＜0.01）,而与巢式PCR差异无统计学意义（P＞0.05）.结论实时PCR检测儿科鼻咽分泌物标本中RSV的方法,敏感性、特异性和可重复性好,可用于急性呼吸道感染患儿标本中的RSV检测.     

北京地区住院急性呼吸道感染患儿的病毒病原检测分析

目的了解北京地区住院急性呼吸道感染(ARI)患儿的病毒病原情况。方法取1260例年龄14岁以下住院ARI患儿的鼻咽深部分泌物,用间接免疫荧光及病毒分离法检测呼吸道合胞病毒(RSV)、流感病毒A、B型、副流感病毒1、2、3型及腺病毒等7种常见呼吸道病毒。用逆转录聚合酶链反应(RT-PCR)法对其中490例患儿标本进行肠道病毒(EV)检测。结果1.ARI1260例中,43.33%检测到了病毒病原,7种常见呼吸道病毒检出率36.19%,RSV阳性率最高(23.97%),以冬春季为著,88.08%的RSV阳性为3岁以下小儿。2.EV阳性率16.33%。3.19例存在2种病毒混合感染,均出现在冬春季,16例为RSV并EV感染。4.入选的510例急性呼气性喘息患儿中,3岁以下RSV阳性率最高(43.20%),3岁以上EV阳性率最高(36.11%)。结论1.RSV是北京地区冬春季婴幼儿ARI的主要病原。2.冬春季EV可并RSV等其他呼吸道病毒感染。3.RSV及EV是引起小儿急性喘息性疾病的主要病毒病原。     

逆转录聚合酶链反应快速检测呼吸道合胞病毒

为探索一种快速、可靠的呼吸道合胞病毒（ＲＳＶ）的检测方法，对逆转录聚合酶链反应（ＲＴ－ＰＣＲ）技术检测ＲＳＶ进行研究。结果：ＲＴ－ＰＣＲ方法可检出病毒量为１０ＴＣＩＤ５０／ｍｌ（ＴＣＩＤ５０为半数组织培养感染量）；用于扩增Ｆ蛋白基因的引物经计算机基因库检测与其他病毒无同源性，实验同步扩增其他常见呼吸道感染病毒均无阳性扩增带；实验过程６小时内完成。在ＲＳＶ好发季节对５３例拟诊为病毒性急性下呼吸道感染婴儿咽拭子进行检测，ＲＳＶ阳性２８例（５２．８％）。提示ＲＴ－ＰＣＲ方法检测ＲＳＶ具有快速、敏感性高、特异性好等特点，适合于临床应用。     

急性下呼吸道感染住院患儿呼吸道合胞病毒感染分析

目的了解急性下呼吸道感染（acutelowerrespiratoryinfection,ALRI）住院患儿呼吸道合胞病毒（respiratorysyncytialviruS,RSV）感染的情况。方法对2010年1月至2013年3月2625例ALRJ住院患儿采用直接免疫荧光法检测其鼻咽分泌物中的RSV抗原。结果送检标本2625例,RSV阳性534例（20．34％）。〈1岁组RSV阳性检出率24．30％（399／1642）,一3岁组15．83％（114／720）,〉3岁组7．98％（21／263）,〈1岁与～3岁组比较（x2=21．102,P〈0．01）,-3岁与〉3岁组比较（X2=10．016,P〈0．01）,差异均有统计学意义。男性患儿RSV阳性检出率21．49％（331／1540）,女性18．71％（203／1085）,男性检出率高于女性（x2=4．579,P〈0．05）。不同年度RSV阳性检出率差异无统计学意义。RSV检出率高峰从每年的10月开始,至次年的4月结束。结论ALRI住院患儿RSV阳性检出率以1岁以内婴儿最高,年龄越小,RSV感染率越高,男性RSV感染高于女性,冬春季为流行高峰。     

应用逆转录—聚合酶链反应鉴别呼吸道合胸病毒亚型

目的 为了使鉴别呼吸道合胞病毒（ｒｅｓｐｉｉｒａｔｏｒｙ ｓｙｎｃｙｔｉａｌ ｖｉｒｕｓ，ＲＳＶ）Ａ、Ｂ亚型的方法更为简单、特异、有效。方法 根据ＲＳＶ Ｇ蛋白编码基因的核苷酸序列设计一套引物，其中ＰＩ为亚型臆通用的引物，Ｐ２和３分别为Ａ、Ｂ亚型特异性引物。将这此引物用于同一逆转录－聚合酶链（ＲＴ－ＰＣＲ）用的引物，Ｐ２和Ｐ３分别为Ａ、Ｂ亚型株的扩增产物分别为２７７ｂｐ和８６３ｂｐ，根据ＰＣＲ产物     

应用逆转录聚合酶链反应鉴别呼吸道合胞病毒亚型

目的为了使鉴别呼吸道合胞病毒（ｒｅｓｐｉｒａｔｏｒｙｓｙｎｃｙｔｉａｌｖｉｒｕｓ，ＲＳＶ）Ａ、Ｂ亚型的方法更为简单、特异、有效。方法根据ＲＳＶＧ蛋白编码基因的核苷酸序列设计一套引物，其中Ｐ１为亚型间通用的引物，Ｐ２和Ｐ３分别为Ａ、Ｂ亚型特异性引物。将这些引物用于同一逆转录聚合酶链（ＲＴＰＣＲ）反应，Ａ、Ｂ亚型株的扩增产物分别为２７７ｂｐ和８６３ｂｐ，根据ＰＣＲ产物的大小即可鉴别所测毒株的亚型。用这一方法对ＲＳＶ原型株和９株我国分离株进行亚型鉴定。结果分离株８株为Ａ亚型，１株为Ｂ亚型，分型结果与单克隆抗体检测和基因序列分析完全相符。结论ＲＴＰＣＲ方法鉴别呼吸道合胞病毒亚型具有快速、简便、敏感、特异等特点，适用于ＲＳＶ临床标本的亚型分型及流行病学研究     

Therapy for respiratory tract infections caused by respiratory syncytial virus

Respiratory syncytial virus (RSV) is the most common viral cause of lower respiratory tract infection (LRTI) in infancy and young children. No effective treatment for RSV lower respiratory tract infection (RSV-LRTI) exists. Ribavirin initially proved to be an effective anti-viral drug for RSV-LTRI. However, subsequently performed trials could not reproduce these positive results and, based on the current available evidence, there is no place for ribavirin in the routine treatment of RSV-LTRI. The use of nebulised bronchodilator therapy in RSV-LTRI has been subject of many trials, with conflicting results. Although the individual patient may have some short-term benefit from nebulised bronchodilators, there does not seem to be a sufficient scientific basis for the standard use of bronchodilator therapy in infants and children with RSV-LTRI. There is increasing evidence that RSV-LTRI is an immune-mediated disease and therefore corticosteroids may be an effective treatment. The results from efficacy trials have demonstrated that corticosteroids are not effective for patients with mild RSV infection. In contrast there are indications that it may be beneficial in patients with more severe RSV-LTRI. It has been demonstrated that in children with RSV infection the vitamin A concentration is inversely related to disease severity. The use of vitamin A in the treatment of patients with RSV-LTRI, however, proved not to be effective. Immunoprophylaxis with hyperimmune immunoglobulins and monoclonal antibody against the viral F-protein have been shown to be effective in the prevention of RSV-LRTI. From the results of the therapeutic efficacy trials, however, it can be discerned that immunoglobulins have no place in the treatment of RSV-LRTI. Conclusion Although respiratory syncytial virus infections each year have a considerable socioeconomic impact, attempts to find an effective therapy have so far been quite unsuccessful. Anti-viral therapy with ribavirin has not been proven to be effective. Symptomatic therapy with bronchodilators may give only short-term relief of symptoms in some individual patients, but has no effect on hospitalisation rates, or duration of hospitalisation. The beneficial effect of corticosteroids in patients with mild respiratory syncytial virus infection is very disappointing, however, there are indications that there might be an effect in patients with more severe infection. So far no beneficial therapeutic effect has been demonstrated with immune globulins. Received: 9 August 1999 / Accepted: 30 November 1999     

2000年冬-2006年春北京地区急性呼吸道感染患儿中呼吸道合胞病毒的监测

目的 对北京急性呼吸道感染患儿进行呼吸道合胞病毒（RSV）的监测并进行亚型分析,探讨其流行规律。方法 2000年11月-2006年3月,于首都儿科研究所附属儿童医院采集因急性呼吸道感染就诊的门诊及住院的患儿咽拭子或鼻咽分泌物标本10048份,接种于Hep-2细胞进行病毒分离,同时用免疫荧光法对鼻咽分泌物标本进行呼吸道病毒抗原的快速检测,对部分RSV阳性的标本用RT-PCR进行了亚型鉴定。结果 （1）在10048份标本中,有2286份为RSV阳性,阳性检出率为22．8％。其中病房标本7176份,RSV阳性2153份,阳性检出率为30．0％;门诊标本2872份,RSV阳性133份,阳性检出率为4．6％。（2）2000～2001年冬春、2002-2003年冬春、2004-2005年冬春RSV的阳性检出率分别为14．0％、18．2％和20．4％,而2001—2002年冬春、2003—2004年冬春和2005-2006年冬春RSV的阳性检出率分别为42．3％、41．0％和40．5％。（3）对938份RSV阳性标本的亚型监测结果：A亚型691份,占73．7％,B亚型247份,占26．3％,2000-2001、2004-2005年冬春季RSV感染以B亚型为主;而2001-2002、2002-2003、2003-2004年冬春季以A亚型为主;2005—2006年为A、B亚型同时流行。结论 RSV是冬春季婴幼儿下呼吸道感染的主要病毒病原,RSV呈现出隔年高峰的流行趋势,RSVA、B亚型是交替出现的,并且有时以相近的比例同时出现。     

小儿呼吸道合胞病毒感染流行特点分析

目的探讨小儿呼吸道合胞感染病毒感染的流行特点及影响RSV流行的气候因素。方法对2001年1月至2006年12月因下呼吸道感染在浙江大学医学院附属儿童医院住院的39597例患儿取鼻咽分泌物标本进行呼吸道合胞病毒(RSV)抗原检测。结果39597例患儿中RSV检测阳性患儿共8657例(21.86%),<1岁者6804例(30.99%),1～3岁者1576例(14.84%),～13岁者277例(3.95%),各年龄组间检出率有统计学意义(P<0.01)。男性5783例(22.76%)高于女性2873例(20.24%),有统计学意义(P<0.01)。RSV检出率与气温之间存在直线负相关,气温每升高1℃,下呼吸道感染患儿中RSV的检出率下降1.8%。冬季是RSV流行高峰,其次是春季和秋季,夏季RSV感染率最低,全年都有RSV的散发流行。结论RSV是冬春季节引起小儿下呼吸道感染的主要病毒,年龄越小RSV感染率越高,男性较女性易感RSV。气温越低,RSV的检出率越高,低气温是杭州地区RSV流行的重要因素之一。     

急性呼吸道感染儿童两亚型呼吸道合胞病毒检测分析

目的 了解2006-2007年度重庆地区住院急性呼吸道感染(ARTIs)儿童两亚型呼吸道合胞病毒(RSV)的感染特点及流行规律.方法 收集2006年4月至2007年3月全年在重庆医科大学附属儿童医院呼吸科住院的部分ARTIs患儿的鼻咽深部吸取物390份,针对RSV G基因保守区序列设计分型引物,采用RT-PCR方法检测标本中RSV的基因组RNA.结果 390例标本中RSV阳性例数为133例(133/390,阳性率为34.10%).阳性标本中A亚型阳性129例,B亚型阳性4例.RSV阳性患儿中,84.9%为2岁以下小儿.2006年11月～2007年1月为RSV高发季节,RSV检测阳性率为55.6%～62.3%,2006年12月RSV检测阳性率最高.B亚型出现于本地区RSV感染低发季节(4、5、6月).RSV感染的临床表现主要为发热(56.4%)、咳嗽(98.5%)、喘息(63.9%)、气促(76.7%)、紫绀(84.9%).临床诊断依次为毛细支气管炎(33.1%),支气管肺炎(27.8%),间质性肺炎(18.1%),重症肺炎伴呼吸衰竭(10.5%),喘息性支气管炎(5.3%),支气管哮喘(4.5%).结论 本研究初步阐明了重庆地区两亚型RSV感染的流行病学特点,证实RSV是重庆地区冬春季婴幼儿ARI的重要病原,2006-2007年度以A亚型RSV流行为主.今后的研究将纳入门诊及社区惠儿并进行多年度连续监测以进一步阐明重庆地区两亚型RSV流行规律及病毒进化特点.     

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.     

下呼吸道感染患儿外周血单个核细胞呼吸道合胞病毒感染的研究

为探讨呼吸道合胞病毒（ＲＳＶ）对外周血单个核细胞（ＰＢＭＣ）的感染情况及感染后细胞免疫的变化，采用免疫组化法对１８例ＲＳＶ性急性下呼吸道感染患儿ＰＢＭＣ内ＲＳＶ及其Ａ、Ｂ亚型抗原进行了检测；采用ＡＰＡＡＰ法、ＭＴＴ比色法和ＥＬＩＳＡ法对Ｔ细胞亚群及Ｔ细胞表面白细胞介素２受体表达、ＰＢＭＣ培养上清液白细胞介素２（ＩＬ－２）活性和可溶性ＩＬ－２受体水平进行了测定。结果显示：１８例ＲＳＶ感染患儿中７例ＰＢＭＣ内可检测到ＲＳＶ抗原；１１例ＲＳＶＡ亚型感染者５例ＰＢＭＣ内均为ＲＳＶＡ亚型阳性，７例ＲＳＶＢ型感染者２例Ｂ亚型阳性；７例恢复期和１０例对照组患儿均为阴性；发病３天以内ＰＢＭＣ中ＲＳＶ抗原阳性者多于３天以后（Ｐ＜０．０５）。ＲＳＶ感染组ＰＢＭＣ内ＲＳＶ抗原阳性者，ＣＤ４细胞比率和ＩＬ－２水平均低于阴性者（ｔ＝２．３８，２．４０，Ｐ值均＜０．０５）。提示：ＲＳＶ性急性下呼吸道感染患儿ＰＢＭＣ可被ＲＳＶ感染，可能由此加重免疫活性细胞损害，导致细胞免疫功能紊乱。     

Burden of respiratory syncytial virus infection in young children

Respiratory syncytial virus (RSV) is the most frequent and important cause of lower respiratory tract infection in infants and children. It is a seasonal virus, with peak rates of infection occurring annually in the cold season in temperate climates, and in the rainy season, as temperatures fall, in tropical climates. High risk groups for severe RSV disease include infants below six mo of age, premature infants with or without chronic lung disease, infants with hemodynamically significant congenital heart disease, infants with immunodeficiency or cystic fibrosis, and infants with neuromuscular diseases. Mortality rates associated with RSV infection are generally low in previous healthy infants (below 1%), but increase significantly in children with underlying chronic conditions and comorbidities. Following early RSV lower respiratory tract infection, some patients experience recurrent episodes of wheezing mimicking early childhood asthma with persistence of lung function abnormalities until adolescence. There is currently no RSV vaccine available, but promising candidate vaccines are in development. Palivizumab, a monoclonal RSV antibody that is the only tool for immunoprophylaxis in high-risk infants, lowers the burden of RSV infection in certain carefully selected patient groups.