直接免疫荧光法对多种呼吸道病毒检测的临床意义

目的为临床提供一种快速诊断呼吸道病毒感染的方法。方法采用直接荧光免疫法检测呼吸道分泌物的合胞病毒、腺病毒、流感病毒A、流感病毒B及副流感病毒1、2、3型。结果对195例呼吸道感染者的鼻咽分泌物标本采用直接荧光免疫法进行病毒原检测,结果显示本地区病毒感染率为57．94％,病毒原以呼吸道合胞病毒为主,副流感病毒1、3型次之,感染以冬季好发。结论该法具有快速、简便、特异性高等优点,对确定临床呼吸道病毒感染是一种非常好的方法。直接荧光免疫法操作简便,费用较低,适合基层医院推广使用。     

Detection of multiple viral agents in nasopharyngeal specimens yielding respiratory syncytial virus (RSV). An assessment of diagnostic strategy and clinical significance

A retrospective study of 6 years (1981-1987) experience with clinical specimens of pediatric patients submitted for identification of respiratory viruses was undertaken to determine the prevalence of multiple viral isolates and to assess the impact of dual infections on severity of clinical disease. Respiratory Syncytial Virus (RSV), the most frequently identified agent, was detected in cell culture and/or by immunofluorescence (IF) in 666 of 2,415 specimens examined. A second virus was isolated in cell cultures from 51 of the 666 specimens (7.6%). Cytomegalovirus, rhinoviruses, adenoviruses, influenza and parainfluenza viruses, echoviruses, vaccine strain polio viruses, and herpes simplex virus were identified with RSV. The diagnosis of a dual viral infection would have been missed in 37 of 51 instances (79%) had rapid diagnosis for RSV been employed without inoculation of cell cultures. Demographics and clinical presentations were similar in patients with dual infections or RSV alone. A case-control study comparing patients with dual isolates and patients with RSV alone to determine the effect of multiple viral infections on severity of disease revealed no significant difference. The combined use of rapid methods and isolation in culture provides more complete viral diagnosis and could have an impact on the choice of antiviral agents and the institution of appropriate infection control measures.     

Comparison of two enzyme-linked immunosorbent assay (EIA) kits with immunofluorescence and isolation in cell culture for detection of respiratory syncytial virus (RSV)

Rapid diagnosis of respiratory syncytial virus (RSV) infections is based upon detection of viral antigen in cells obtained from the respiratory tract and usually employs immunofluorescence (IF) reactions or enzyme-linked immunosorbent assays (EIA). The Pathfinder EIA kit (Kallestad Diagnostics) was compared with the Abbott EIA kit by evaluating each against isolation of RSV in cell culture and detection of antigen by IF. The Pathfinder kit identified 116 of 129 culture-positive and 72 of 90 culture-negative specimens; the sensitivity was 90 percent and the specificity was 80 percent. The sensitivity of the Abbott EIA test compared to isolation of RSV in cell culture was 91% (115 of 127), and the specificity was 83% (74 of 89). Of 165 specimens evaluated by IF, the Pathfinder kit detected 97 of 105 IF-positive and 45 of 60 IF-negative specimens, giving a sensitivity of 92% and a specificity of 75%. The Abbott EIA compared similarly with IF, showing a sensitivity of 91% (98 of 108) and a specificity (42 of 54) of 78%. Visual reading of the Kallestad test resulted in a sensitivity of 92%, a specificity of 91%, positive predictive value of 95%, and negative predictive value of 86%. The Pathfinder EIA kit compared well with IF and the Abbott EIA for detection of RSV antigen but performed faster than the Abbott test and offers the option of a visual reading.     

海岛地区婴幼儿呼吸道感染的病原学研究

目的 探讨海岛地区婴幼儿急性呼吸道感染的病毒病原.方法 采取211例急性呼吸道感染患儿的痰液做病毒检测.结果 病毒感染35例,其中呼吸道合胞病毒(RSV)感染20例(9.5%),流感病毒感染6例(2.8%),腺病毒9例(4-3%),副流感病毒Ⅲ型4例(1.9%),副流感病毒Ⅰ、Ⅱ型各2例(1%).结论 RSV感染仍是海岛婴幼儿急性呼吸道病毒感染最常见的病原,病毒病原的诊断可为临床诊断和治疗提供可靠的依据.     

Anti-RNA virus activity of polyoxometalates.

The anti-RNA virus activity of polyoxometalates (POM) is reviewed, with a special emphasis on the anti-respiratory virus activities. There are many causative agents of acute viral respiratory infections; and it is rather difficult to identify the relevant agent in a given case by rapid clinical means. During acute progress of infection before the definitive diagnosis is obtained physicians need to prescribe certain broad spectrum anti-viral drugs. A titanium containing polyoxotungstate, PM-523 exhibited potent anti-influenza virus (FluV) A and anti-respiratory syncytial virus (RSV) activities in vitro. Therapeutic effect of FluV A infected mice with aerosol inhalation of PM-523 was proven. A vanadium substituted polyoxotungstate, PM-1001 has antiviral activity against FluV A, RSV, parainfluenza virus (PfluV) type 2, Dengue fiver virus, HIV-1 and SARS coronavirus in vitro. Thus, POMs have been proven to be broad spectrum and non-toxic anti-RNA virus agents in both in vitro and in vivo experiments and are promising candidates for first-line therapeutics in acute respiratory diseases.     

2016-2020年贵阳市老年呼吸道感染住院患者呼吸道合胞病毒流行特征及流感病毒感染临床严重性比较

目的 了解老年人呼吸道感染住院患者呼吸道合胞病毒(RSV)感染的流行病学特征,比较RSV感染与流感病毒感染的临床严重性差异,探索影响老年人呼吸道感染住院期间死亡的危险因素。方法 收集2016年1月1日至2020年12月31日贵阳市2家医院收治的60岁以上老年人呼吸道感染住院患者的临床和病原学检测资料。比较RSV感染与流感病毒感染老年人在人口学特征、基础性疾病、临床表现,以及住院时间和住院期间死亡等临床结局上存在的差异,并采用多因素logistic回归模型探索影响老年人呼吸道感染住院期间死亡的危险因素。结果 5 131例60岁以上呼吸道感染住院患者中,RSV检测阳性率为2%,流感病毒检测阳性率为10%。77%(80/104)的RSV感染和63%(312/497)的流感病毒感染集中在每年的11月至次年2月,发病具有明显季节性。与流感病毒感染老年人比较,RSV感染的中位发病年龄(73岁vs. 74岁,P=0.997),ICU收治比例(7%vs. 6%,P=0.822)和住院期间死亡率(6%vs. 3%,P=0.233)差异无统计学意义。但RSV感染的老年人中位住院天数延长(14 d vs. ...     

Monoclonal antibodies for the rapid diagnosis of respiratory syncytial virus infection by immunofluorescence

Eight separate monoclonal antibodies to the Long strain of respiratory syncytial virus (RSV) were tested for their utility as rapid diagnostic reagents in immunofluorescence. Preliminary screening indicated that all 8 reacted with 11 field strains from three previous local RSV outbreaks and with 4 of 5 additional strains chosen because of their antigenic diversity by neutralization. Two monoclonal antibodies, one each directed against a surface glycoprotein and the nucleocapsid protein, were then compared, singly and combined, with a polyclonal antiserum as diagnostic reagents in 209 consecutive samples submitted to our diagnostic laboratory. Agreement between the two monoclonal antibodies was 100% and between them and the polyclonal serum was 98%. Sensitivity in relation to culture was 96–;98%. Monoclonal antibodies are excellent immunofluorescent diagnostic reagents; antigenic diversity among RSV strains was not an impediment to their use in this study.     

Enzyme immunoassay for respiratory syncytial virus: Rapid detection in nasopharyngeal secretions and evaluation of isolates representing different RSV subgroups

The presence of respiratory syncytial virus (RSV) was investigated by immunofluorescent antibody (IFA) technique and by an enzyme immunoassay (EIA) in 169 samples of nasopharyngeal secretions of infants and children with acute respiratory infections. Of 31 samples positive by EIA, 25 were positive by IFA. In 24 samples from a retrospective study, RSV positive by IFA and/or tissue culture isolation (TCI), 22 were also positive by EIA. The EIA was also evaluated with 111 RSV isolates in Hep2 cell cultures representing different RSV subgroups. All were positive by EIA. © 1993 Wiley-Liss, Inc.     

Changing patterns of detection of viruses in respiratory specimens since introduction of monoclonal antibody-based immunofluorescence test for respiratory syncytial virus antigen

In our earlier studies, a rapid monoclonal antibody-based direct immunofluorescence (IF) test for respiratory syncytial virus (RSV) antigen in nasopharyngeal aspirates (NPAs) proved sensitive and specific, and dual virus infections were very rare when the test was used during the winter RSV season. We therefore recommended that virus isolation in cell culture be restricted to RSV antigen-negative specimens when RSV infection was highly prevalent (October to March in the UK). We reviewed the effect of this policy on the detection of RSV and other viruses in NPAs. The number of NPAs received each RSV epidemic increased markedly with time (3804 in 1984–88, 4569 in 1989–93, P < 0.0001), while during the test of the year the numbers declined slightly (753 vs. 700 for the same periods, P < 0.05). The RSV positivity rate increased October to March (1447/3588 (40%) in 1984–89, 2357/4591 (48%) in 1989–94, P < 0.0001). Restricted use of virus isolation led to a dramatic decline in the yield of RSV in cell culture from RSV IF-negative NPAs (196/3804 (5.2%) for 1984–88, 30/4569 (0.66%) for 1989–93, P < 0.0001). The already low yield on non-RSV respiratory viruses in cell culture was not affected by this policy. A strategy is proposed for the detection of RSV and other respiratory viruses in NPAs which may make virus isolation on such specimens obsolete.     

Surveillance of viral respiratory tract infections over a one year period in mainly hospitalized Austrian infants and children by a rapid enzyme-linked immunosorbent assay diagnosis

In order to obtain more information on viral respiratory tract infections in Austrian infants and children, nasopharyngeal secretions from 1432 infants and children, collected from October 1984 to October 1985, were screened for the presence of respiratory syncytial virus (RSV), adenoviruses, parainfluenza virus type 1, 2, and 3, and influenza viruses type A and B, by enzyme-linked immunosorbent assay (ELISA). The results obtained were analyzed with respect to incidence, seasonal distribution and clinical syndromes associated with the different viral pathogens investigated and also with the practicability of ELISA diagnostics over long distances. A viral etiology of acute respiratory tract infection was confirmed in 372 (26%) infants. RSV was detected in 286 (20%) of the nasal secretions and was thus the most frequently encountered agent. RSV infections occurred mainly in the winter months and were often associated with bronchitis, bronchiolitis, and pneumonia. Only sporadic infections were found with one of the other viruses investigated.     

Viral infections of the respiratory tract in patients with cystic fibrosis

The role of viruses and bacteria in the development of respiratory tract infections causing acute deteriorations in lung function in patients with cystic fibrosis (CF) was investigated. Over a period of 30 months, 29 viral respiratory diseases were proven serologically by testing 275 sporadically collected sera from 75 patients with cystic fibrosis. The influenza A virus was the most frequent responsible viral pathogen (11 × ), followed by adenovirus (8 × ), influenza B virus (5 × ), parainfluenza virus type 3 (3 × ), parainfluenza virus type 1 and respiratory syncytial virus (RSV) (each of 1 × ). There was no serological evidence for infections with parainfluenza virus type 2, Mycoplasma pneumoniae or Coxiella burnetii. Deterioration of the clinical condition was found in 78% of the viral infections leading in 70% to hospital admission. Patients with cystic fibrosis and viral respiratory illnesses showed significantly more admissions to the hospital (3·2 ± 2·7) with a longer stay (90·6 ± 99·6 days). Nearly all viral episodes (93%) were accompanied or followed by a significant change of the microbial flora in the sputum especially by colonisation with Pseudomonas aeruginosa, Staphylococcus aureus and Haemophilus influenzae. Seventy-two per cent of the viral infections occurred at home and 28% seemed to be hospital acquired. Our study emphasises the importance of improving antibacterial therapy at home to reduce the number of hospital admissions. Efforts for prophylaxis by vaccination or the use of chemotherapeutic agents should be made for the patients with cystic fibrosis.     

常德地区小儿呼吸道病毒性感染病原学分析

目的探讨常德地区小儿呼吸道病毒性感染的病原学概况,为临床诊治工作提供参考.方法应用免疫荧光法检测患儿早期咽拭子标本中7项病毒[合胞病毒(RSV)、腺病毒(ADV )、流感病毒 A (FLuA )、流感病毒B(FLuB)、副流感病毒1(PIV1)、副流感病毒2(PIV2)、副流感病毒3(PIV3)],以了解病原学分布.结果从2778例患儿咽拭子标本中,检出相关病毒阳性1260例,总阳性率为45.4%.含单项病毒感染占89.0%,2项混合感染占11.0%.单一病毒感染中 RSV 占首位(57.40%),其余依次为 PIV1(11.31%)、PIV2(8.91%)、FLuB(6.33%)、ADV(5.97%)、PIV3(5.53%)、FLuA(4.55%).结论RSV 、PIV1、PIV2为常德地区致小儿呼吸道感染的主要流行病原体.对疑似病例早期进行7项呼吸道病毒检测,尽早为临床提供病原学诊断,使患儿得到及时诊治.     

2019年湖北省武汉市儿童呼吸道合胞病毒感染临床特征及流行病学分析

目的分析2019年湖北省武汉市呼吸道合胞病毒(RSV)引起儿童急性呼吸道感染(ARTI)的流行病学特征,总结其临床特点。方法收集2019年因ARTI在武汉儿童医院治疗的患儿鼻咽拭子标本。采用直接免疫荧光法检测样本中的RSV,根据临床资料进行流行病学分析。结果 2019年武汉市ARTI患儿中RSV阳性率为8.41%,不同年龄段患儿检出率差异有统计学意义(P<0.05),且RSV的检出率随患儿年龄增加而逐渐降低。男性患儿中的检出率高于女性患儿(P<0.05)。RSV在1-2、12月检出率最高,在5-7月检出率最低。患儿临床表现主要为咳嗽、发热、喘息。部分患儿可出现其他并发症,如肝功能异常、癫痫、心律失常、中耳炎等。RSV与其他呼吸道病毒的混合感染率为2.81%,主要为副流感病毒3型及腺病毒。结论 RSV是武汉市儿童ARTI的常见病原,流行时间为12月至次年2月,患儿年龄越小越容易被感染。RSV易与副流感病毒3型及腺病毒发生混合感染。     

珠海地区353例儿童喘息性疾病病毒病原学分析

目的 探讨儿童喘息性疾病的病毒病原学特征,提高临床诊治水平.方法 收集本院儿科2018年6月至2019年5月353例喘息患儿的咽拭子,应用多重PCR技术对病毒进行检测,并分析临床资料.结果 病毒检测阳性124例(35.13％),呼吸道合胞病毒(RSV)77例(21.81％)、副流感病毒(PIV)14例(3.97％)、博...     

Epidemiological aspects of respiratory syncytial virus (RSV) infections in Italy: a national survey

Respiratory syncytial virus (RSV) is a major cause of respiratory infections such as bronchiolitis and pneumonia in babies and children, producing serious infections and short- to medium-term complications. We conducted a survey, using a questionnaire, to determine the opinions of paediatricians on the geographical and seasonal distribution, and the clinical impact of RSV in Italy. Of the 344 paediatricians who responded, 66% had admitted children affected by RSV into their department, and 38% of these had admitted more than 10 cases during the preceding year. According to those interviewed, newborn babies of lowest gestational age were most often affected. The children considered to be at highest risk of severe infection were those with bronchodysplasia, immunodeficiencies or congenital heart disease; most admissions (87%) were attributed to bronchiolitis. In southern Italy and the islands RSV appeared later and persisted later into the year than in central and northern Italy. Most paediatricians (79%) considered the available treatments for serious RSV ineffective but only half were aware of the possibility of immunoprophylaxis against RSV.     

A colorimetric microtiter plate PCR system detects respiratory syncytial virus in nasal aspirates and discriminates subtypes A and B.

We developed a colorimetric microtiter plate (MTP) PCR system for specific detection of the respiratory syncytial virus (RSV) nucleocapsid gene and differentiation of viral subtypes A and B. Of 47 pediatric nasal aspirate specimens, the sensitivity and specificity were 94.4% (17 of 18) and 100% (15 of 15), respectively, when compared with RSV cell culture isolation in HEp-2 cells. An additional 14 specimens positive for adenoviruses, rhinoviruses, influenza, or parainfluenza viruses did not give positive reactions. PCR testing detected a mean of 0.15 (0.01 to 7.00) plaque-forming units of RSV virions. Inhibition of PCR amplification was detected in 33.3% (6/18) of undiluted specimens and could be avoided by a dilution (1:10) of extracted RNA without decreasing test sensitivity. RSV subtype, as determined by allele-specific probes, was identical to that determined by an immunofluorescence assay. These results indicate that the MTP PCR system provides a sensitive and specific test for clinical laboratory diagnosis and simultaneous subgroup classification of RSV infection.     

福州地区急性呼吸道感染患儿呼吸道合胞病毒检测及亚型分析

目的:分析福州地区2006-2007年冬春季我院收治急性呼吸道感染患儿呼吸道合胞病毒(RSV)及其亚型流行情况。方法:采用RT-PCR方法检测176例急性呼吸道感染患儿的鼻咽拭子中RSV并进行亚型鉴定分析。结果:(1)在176例标本中,RSV阳性28例(15.9%)。其中急性下呼吸道感染阳性率为38%,显著高于急性上呼吸道感染的4%(!2=33.15,P<0.05)。(2)28例RSV阳性标本中,A亚型23例(82%),B亚型4例(14%),不明分型者1例(4%);上、下呼吸道感染组患儿间RSV亚型分布差异无显著性(P>0.05),都以A型为主。(3)RSV阳性患儿的年龄主要以3岁以下为主。结论:RSV是婴幼儿急性下呼吸道感染的重要病原体;福州地区2006-2007年冬春季RSV两种亚型同时流行,以A亚型为主。     

Respiratory syncytial virus infection in children

Respiratory syncytial virus (RSV) is an RNA virus that causes respiratory tract infections in children. In the North- ern Hemisphere, the peak infection season is November through April. By two years of age, most children will have had an RSV infection. Bronchiolitis, a lower respiratory tract infection, is often caused by RSV. An RSV infection is diagnosed based on patient history and physical examination. Children typically present with cough, coryza, and wheezing. Laboratory testing and chest radiography are not necessary to make the diagnosis. Serious concur- rent bacterial infections are rare. Treatment of an RSV infection is supportive, with particular attention to maintaining hydration and oxygenation. Children younger than 60 days and those with severe symptoms may require hospitalization. Neither antibiotics nor corticosteroids are helpful for bronchiolitis. A bronchodilator trial is appropriate for children with wheezing, but should not be continued unless there is a prompt favorable response. Frequent hand washing and contact isolation may prevent the spread of RSV infections. Children younger than two years at high risk of severe illness, including those born before 35 weeks of gestation and those with chronic lung or cardiac problems, may be candidates for palivizumab prophylaxis for RSV infection during the peak infection season. Most children recover uneventfully with supportive care.