This study aimed to develop a portable, accurate and easy-to-operate scheme for rapid detection of respiratory virus nucleic acid. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to verify the effect of extraction-free respiratory virus treatment reagent (RTU) on viral nucleic acid treatment and the effect of ultra-fast fluorescence quantitative PCR instrument (FQ-8A) on nucleic acid amplification, respectively. RTU and FQ-8A were combined to develop a rapid detection scheme for respiratory virus nucleic acid, and the positive detection rate was judged by C_t value using a fluorescence quantitative PCR instrument, and the accuracy of the scheme in clinical samples detection was investigated. The results showed that RTU had comparable sensitivity to the automatic nucleic acid extraction instrument, its extraction efficiency was comparable to the other 3 extraction methods when extracting samples of different virus types, but the extraction time of RTU was less than 5 min. FQ-8A had good consistency in detection respiratory syncytial virus (RSV) and adenovirus (ADV) compared with the control instrument ABI-7500, with kappa coefficients of 0.938 (P < 0.001) and 0.887 (P < 0.001), respectively, but the amplification time was only about 0.5 h. The RTU and FQ-8A combined rapid detection scheme had a highly consistent detection rate with the conventional detection scheme, with a sensitivity of 91.70% and specificity of 100%, and a kappa coefficient was 0.944 (P < 0.001). In conclusion, by combining RTU with FQ-8A, a rapid respiratory virus nucleic acid detection scheme was developed, the whole process could be completed in 35 min. The scheme is accurate and easy-to-operate, and can provide important support for the rapid diagnosis and treatment of respiratory virus.
Humans ; Respiratory Syncytial Virus Infections/diagnosis* ; Respiratory Syncytial Virus, Human/genetics* ; Nucleic Acid Amplification Techniques ; Real-Time Polymerase Chain Reaction ; Adenoviridae ; Sensitivity and Specificity

OBJECTIVETo characterize the prevalence and occurrence of subgroups of human respiratory syncytial virus (RSV) in infants and young children with acute respiratory infections (ARI) in Beijing area.

METHODSRSVs were identified from nasopharyngeal aspirates and throat swabs collected from infants and children with ARI who visited the Children's Hospital Affiliated to Capital Institute of Pediatrics during the period of November 2000 to March 2006, by virus isolation in Hep-2 cells and antigen detection by indirect immunofluorescence assay (IFA). RT-PCR was used to differentiate subgroups A and B of RSV from part of the positive specimens.

RESULTSOut of 10 048 specimens including 7176 nasopharyngeal aspirates from inpatients and 2872 throat swabs from outpatients, 2286 (22.8%) were RSV positive. The positive rate for RSV identification were 30.0% (2153/7176) in specimens from the hospitalized patients, which was higher than that from outpatients (4.6%, 133/2872). The youngest of the RSV positive patients was 1 day after birth and the oldest was 15 years of age, with 73.0% younger than 1 year. Among those RSV positives, only 1.6% were older than 5 years. The ratio of male to female who were RSV positive was 2.4:1 (1598:674). The clinical diagnosis for 91.2% (1991) of those RSV positive patients was severe lower respiratory infections including bronchiolitis and pneumonia, whereas in only 8.8% (192) the diagnosis was upper respiratory infections. The data revealed that RSV started to be detected in October each year during the survey period and November to next April was the RSV season. The detection rate declined in May and almost no RSV could be found in summer. Positive rates for RSV detection were 42.3%, 41.0% and 40.5% in the seasons of 2001 - 2002, 2003 - 2004, 2005 - 2006, which were higher than those in seasons of 2000 - 2001 (14.0%), 2002 - 2003 (18.2%), 2004 - 2005 (20.4%). Subtyping of A and B during the surveillance period showed that 73.7% (691/938) were subgroup A and 26.3% (247/938) were subgroup B. Subgroup B was predominant in the 2000 - 2001 and 2004 - 2005 seasons, whereas subgroup A predominated in the 2001 - 2002, 2002 - 2003 and 2003 - 2004 seasons. Almost equal proportions of subgroup A and B appeared in 2005 - 2006 seasons.

CONCLUSIONThe data indicate that RSV is an important etiological agent for lower respiratory infections in infants and young children in winter and spring during the survey period. The pattern of RSV circulation varied alternately with higher rate every other year. The predominant subgroup changed between A and B, and co-circulated in equal proportion in some years.

Adolescent ; Cell Line, Tumor ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Population Surveillance ; Prevalence ; Respiratory Syncytial Virus Infections ; diagnosis ; epidemiology ; Respiratory Syncytial Virus, Human ; genetics ; isolation & purification ; Respiratory Tract Infections ; diagnosis ; epidemiology ; virology ; Seasons

OBJECTIVETo study the application of association rules in Chinese medical pathogeneses and pathologies of heat and phlegm in infantile viral pneumonia.

METHODSAssociation rules were applied to analyze dynamic changes of heat and phlegm correlated symptoms and signs in 297 infants with respiratory syncytial virus (RSV) pneumonia, thus understanding its evolution or pathogenesis.

RESULTSHeat and phlegm co-exist in infantile viral pneumonia. In their relationship, heat was more likely to affect phlegm, but phlegm was less likely to affect heat. Under the intervention of drugs, the possibility of heat induced by phlegm was gradually reduced. But the possibility of phlegm induced by heat was not obvious as time went by.

CONCLUSIONSHeat and phlegm have a close relationship in the pathogenesis of infantile viral pneumonia. The intervention of drugs could reduce the pathologic evolution of phlegm causing heat. However, it has little effect on the pathologic evolution of heat causing phlegm.

Child, Preschool ; Female ; Humans ; Infant ; Male ; Medicine, Chinese Traditional ; methods ; Pneumonia, Viral ; diagnosis ; Respiratory Syncytial Virus Infections ; diagnosis

Child, Preschool ; Humans ; Infant ; Infant, Newborn ; Lung ; physiopathology ; virology ; Male ; Metapneumovirus ; isolation & purification ; Paramyxoviridae Infections ; diagnosis ; physiopathology ; Respiratory Function Tests ; Respiratory Syncytial Virus Infections ; diagnosis ; physiopathology ; Respiratory Syncytial Viruses ; isolation & purification

OBJECTIVETo understand the clinical characteristics and outcome associated with respiratory syncytial virus (RSV) infection in hematopoietic stem cell transplantation (HSCT) recipients with primary immunodeficiencies (PIDs).

METHODNasopharyngeal aspirate samples were collected consecutively before and after HSCT from 9 recipients from Apr. 2009 to Sep. 2010 and analyzed for the presence of RSV using real-time polymerase chain reaction assay. To further verify the presence of the virus, positive samples for PCR were isolated for RSV. RSV G gene was amplified, sequenced and used for phylogenetic analysis.

RESULTThe presence of RSV was detected in 3 out of 9 children. The viral replication in all the patients was prolonged for months. All the 3 patients with RSV infection were treated with intravenous immune globulin (IVIG) and one was treated with antiviral medication. All patients survived and achieved successful immune reconstitution.

CONCLUSIONThis study indicates that the HSCT recipients with PID are at increased risk for RSV infection. RSV can shed for months after the initial infection and the patients recover with the course of immune reconstitution.

Child, Preschool ; Female ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Immunologic Deficiency Syndromes ; surgery ; virology ; Infant ; Prognosis ; Respiratory Syncytial Virus Infections ; diagnosis ; drug therapy ; Respiratory Syncytial Viruses ; genetics ; isolation & purification ; physiology ; Virus Replication ; Virus Shedding

OBJECTIVETo establish a rapid, sensitive and specific reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for detecting human respiratory syncytial virus (RSV) in respiratory samples collected from children with acute respiratory infections.

METHODAccording to the conserved matrix gene sequences of respiratory syncytial virus subtypes A and B downloaded from GenBank, primers were designed and RT-LAMP assay was developed to detect RNA of RSV sensitivity of the RT-LAMP method was evaluated by using ten-fold serially diluted in vitro-transcribed matrix RNA fragments from RSV A and RSV B, respectively. Specificity of the RT-LAMP method was tested through cross-reaction with other RNA and DNA viruses. Then 5 RSV strains isolated from clinical specimens using tissue cultures were tested by RT-LAMP assay. A total of 101 nasopharyngeal aspirates from hospitalized patients with acute respiratory infections which had been tested by direct immunofluorescence assay (DFA), including 40 positive for RSV and 61 negative for RSV, were tested by RT-LAMP assay and by RT-nested PCR.

RESULTSensitivity analysis indicated that this RT-LAMP method was able to detect 1 copy/µl of RSV A and RSV B RNA, no amplification was shown in RT-LAMP with DNA or cDNA from other viruses in 60 min, revealed that the RT-LAMP assay is highly specific. Five RSV isolates confirmed as 4 RSV A and 1 RSV B previously were detected by RT-LAMP method as positive in 30 min. For those 101 specimens tested, 37 were RSV positive determined by RT-LAMP assay, as well as 35 RSV positive by RT-nested PCR. The total coincidence rate of RT-LAMP assay with DFA and RT-nested PCR in detecting RSV is 95.0%, 94.1% with Kappa value 0.895 and 0.871, respectively.

CONCLUSIONA new, sensitive, accurate and rapid method, RT-LAMP assay for detecting human respiratory syncytial viruses from nasopharyngeal aspirates was developed, which should be helpful in rapid detection of RSV from respiratory tract samples of children.

Acute Disease ; Child ; Child, Preschool ; DNA Primers ; Humans ; Infant ; Molecular Diagnostic Techniques ; Nasopharynx ; virology ; Nucleic Acid Amplification Techniques ; RNA, Viral ; isolation & purification ; Respiratory Syncytial Virus Infections ; diagnosis ; Respiratory Syncytial Virus, Human ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity

OBJECTIVEThe impact of human bocavirus (HBoV), a newly identified human parvovirus, on childhood persistent wheezing has not been identified. In this study, the clinical features of infantile persistent wheezing induced by HBoV was analyzed.

METHODSTracheal aspirates were collected by bronchofibroscope or nasopharyngeal (NP) aspirates from April, 2006 to January, 2007. HBoV DNA in the tracheal aspirates of 33 children with persistent wheezing and in NP aspirates of 6 children with persistent wheezing, who had at least or more than four weeks wheezing. RSV was identified by virus isolation in Hep-2 cells and antigen detetion by direct immunofluorescence assay (DIFA) which was also used for diagnosis of adenovirus, influenza A and B, parainfluenza 1, 2, 3 infection.

RESULTSOf the 39 children with persistent wheezing, 12 cases (31%) were positive for HBoV DNA. Age of HBoV-positive patients ranged from 2 month to 1 year. The results of sequencing of PCR products proved that sequences of HBoV DNA from these 12 samples were exactly identical to the those of HBoV stored in GeneBank (accession numbers DQ000495 and DQ000496). Two cases with HBoV infection were found to be co-infected with RSV. Ten of the 12 HBoV-positive samples were collected during the period from winter to spring (1 in November, 4 in December, 2 in January and 3 in April), the other two HBoV-positive samples were collected during the period from summer to autumn (1 in May and the other in July). Seven of the 12 HBoV DNA-positive patients had fever, 5 of them had high fever. Significantly more patients with HBoV infection had fever as compared to patients with RSV infection. All the HBoV positive patients showed abnormal findings on chest X ray such as interstitial infiltrates, lung infiltration and hyperinflation. Abnormal findings on chest X ray were found in higher proportion of HBoV positive patients as compared with RSV positive patients. And other manifestations such as wheezing, cough and respiratory distress had no significant difference between HBoV and RSV infected patients.

CONCLUSIONSThis study further demonstrated that HBoV probably is a common pathogen of lower respiratory infection in children and might particularly be associated with persistent wheezing.

Child, Preschool ; Cough ; etiology ; Female ; Fever ; etiology ; Human bocavirus ; pathogenicity ; Humans ; Infant ; Male ; Nasopharynx ; pathology ; Paramyxoviridae Infections ; physiopathology ; Parvoviridae Infections ; physiopathology ; Respiratory Sounds ; etiology ; Respiratory Syncytial Virus Infections ; classification ; physiopathology ; Respiratory Tract Infections ; diagnosis ; physiopathology ; virology

OBJECTIVEThe present study was designed to explore the practical application of the rapid etiological diagnosis by detecting specific IgM antibody against common respiratory viruses in children with acute lower respiratory infections (ALRI).

METHODClinical specimens including nasopharyngeal aspirates and serum of acute phase from hospitalized children were collected from 207 infants and children with acute lower respiratory infections from March 2009 to September 2010. Seven common respiratory virus antigens were identified from the collected nasopharyngeal aspirates by direct immunofluorescence assay (DFA). ELISA was used to detect specific IgM antibody against RSV, ADV, IFVA, IFVB and PIV, while indirect immunofluorescence assay (IFA) was used to detect specific IgM antibody against RSV, ADV, IFVA, IFVB, PIV1, PIV2 and PIV3 in collected acute phase serum.

RESULTThe overall positive rates to detect viral antigen by using DFA, ELISA and IFA was 67.6%, 57.5% and 39.6%, respectively. The consistent rate of ELISA and IFA versus accepted DFA were 21.7% and 31.4%, respectively. The average days from onset of the symptoms to blood sample collection for those with the consistent results by ELISA and DFA were 12.0 d for ADV, 9.6 d for PIV2, 9.5 d for IFV, and 5.3 d for RSV, respectively, and by IFA and DFA were 15.0 d for PIV3, 9.2 d for ADV, and 7.4 d for RSV, respectively. Among all age groups, the consistent rate of serum viral IgM and antigen detections was highest in children younger than 3 years old.

CONCLUSIONAlthough there were differences between serum IgM antibody and viral antigen detections, specific IgM antibody detection was of value in early and rapid etiological diagnosis of pediatric ALRI, especially for young children. It could provide serologic evidence of respiratory virus infection. The diagnostic rate of pathogen could be improved if it was used in combination with viral antigen diagnostic methods.

Antibodies, Viral ; analysis ; blood ; Antibody Specificity ; Antigens, Viral ; analysis ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Female ; Fluorescent Antibody Technique ; Humans ; Immunoglobulin M ; analysis ; blood ; Infant ; Male ; Nasopharynx ; virology ; RNA Viruses ; genetics ; isolation & purification ; Respiratory Syncytial Virus Infections ; diagnosis ; virology ; Respiratory Syncytial Viruses ; genetics ; isolation & purification ; Respiratory Tract Infections ; diagnosis ; immunology ; virology ; Sensitivity and Specificity

This study was performed to evaluate the utilization and outcomes of palivizumab in high risk children born prematurely with chronic lung disease (CLD). A retrospective review of 128 patients was conducted from September 2004 to March 2009 at the Ajou University Hospital. All patients were diagnosed with CLD, were born at < or =35 weeks of gestation, were <2 yr old at the onset of respiratory syncytial virus (RSV) season, and had received medical therapy within six months prior to the RSV season. Fifty-three patients did not receive palivizumab prophylaxis and 75 patients received at least one dose of palivizumab. There were no statistically significant differences between the patients with and without palivizumab prophylaxis with regard to demographic characteristics and risk factors for RSV infection. There were no systemic adverse responses. Compliance with the course of prophylaxis was 92.2%. Hospitalization associated with RSV occurred in 12 cases (22.6%) in the group without prophylaxis and in three cases (4.0%) with prophylaxis. Palivizumab prophylaxis significantly reduced the frequency of RSV-related hospitalization in preterm children with CLD. This is the first retrospective review of palivizumab prophylaxis in Korea. Palivizumab is effective and well tolerated in high risk prematurely born children.
Antibodies, Monoclonal/*therapeutic use ; Antibodies, Monoclonal, Humanized ; Antiviral Agents/*therapeutic use ; Bronchopulmonary Dysplasia/*complications/diagnosis ; Demography ; Hospitalization ; Humans ; Infant ; Infant, Newborn ; Premature Birth ; Republic of Korea ; Respiratory Syncytial Virus Infections/complications/*prevention & control ; Retrospective Studies ; Risk Factors ; Treatment Outcome

OBJECTIVETo evaluate the curative effect of Qingkailing Injection (QKL) for treatment of children's respiratory syncytial virus pneumonia of phlegm-heat obstructing Fei syndrome pattern (SVP-PH) depending upon main symptom assessment.

METHODSA Chinese-Western medicine comparative trial was conducted on 206 children with SVP-PH in two groups treated with Ribavirin injection plus compound guaiacol potassium sul-fonale oral solution (as control group) and QKL injection plus Ertong Qingfei oral liquid (as treated group) respectively, for 10 days. The curative effectiveness on four main symptoms (fever, cough, sputum and short breath) were evaluated at different time-points.

RESULTSThe effectiveness in the treated group at various time-points was superior to that in the control group, showing the earlier initiating time (on the 4th day) and the preponderances on cough and sputum ran all through the whole course.

CONCLUSIONChinese medicine shows a multi-target effect in treating children's SVP-HP.

Child ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Guaiacol ; analogs & derivatives ; therapeutic use ; Humans ; Infusions, Intravenous ; Male ; Medicine, Chinese Traditional ; Phytotherapy ; Pneumonia, Viral ; drug therapy ; Respiratory Syncytial Virus Infections ; drug therapy ; Ribavirin ; therapeutic use