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Use of a thin film biosensor for rapid visual detection of PCR products in a multiplex format.

Rapid, sensitive assays for nucleic acid amplification products have utility for the identification of bacterial or viral infections. We have developed a nucleic acid hybridization assay utilizing thin film technology that permits visual detection of hybrids. The silicon-based biosensor detects the presence of target sequences by enzymatically transducing the formation of nucleic acid hybrids into molecular thin films. These films alter the interference pattern of light on the biosensor surface, producing a perceived color change. We have applied this technology to the development of a chip containing capture probes specific for human respiratory virus sequences including respiratory syncytial virus, influenza virus A and B, parainfluenza virus types 1 and 3, and rhinovirus. In a ten-minute assay, the biosensor permits unambiguous identification of viral-specific RT/PCR products from infected cell lysates.

Base Sequence↗

Nosocomial viral infections: I. Epidemiology and significance.

Viral illnesses in Strong Memorial Hospital were monitored over a 17-month period. Using criteria based primarily on the incubation periods for a number of common virus infections, the infections we found were classified as hospital- or community-acquired. Hospital-acquired viral infections occurred on most hospital services; the majority of infections occurred on the pediatric and psychiatric services. Infections due to herpesviruses were seen more frequently in a group of patients aged 14 years or older, while infections in patients aged three years or younger were more likely to be due to respiratory syncytial virus, influenzavirus, adenovirus, or parainfluenza virus. Patients with nosocomial infections due to viruses were hospitalized an average of 9.3 days longer than uninfected controls; thus nosocomial viral infections result in increased costs of hospitalization.

Adenoviridae Infections↗

Respiratory virus infections in stem cell transplant patients: the European experience.

The frequency of and survival from community-acquired respiratory virus (CRV) infections among patients undergoing allogeneic or autologous stem cell transplantation (SCT) were evaluated in a prospective study conducted at 37 medical centers affiliated with the European Group for Blood and Marrow Transplantation. Of the 40 CRV infections diagnosed in 1863 patients (739, allogeneic SCT; 1124, autologous SCT), 20 were attributed to respiratory syncytial virus (RSV), 4 to parainfluenza viruses, and 16 to influenza virus A. The overall survival rate among SCT recipients with CRV infections was 76%; 8 patients, all recipients of allogeneic transplants, died after diagnosis of CRV infection, but only 5 of these deaths (3, RSV; 2, influenza A virus) were attributable to the infection. The overall rate of death directly attributable to RSV and influenza A virus infections in allogeneic SCT recipients was 1.1%. In an 18-month extension, an additional 53 patients with CRV were identified. Results for the combined data were similar to those from the first phase of the study.

Antiviral Agents↗

The association of newly identified respiratory viruses with lower respiratory tract infections in Korean children, 2000-2005.

BACKGROUND: This study was performed to evaluate the associations of newly recognized viruses, namely, human metapneumovirus (hMPV), human coronavirus (HCoV)-NL63, and human bocavirus (HBoV) with lower respiratory tract infections (LRTIs) in previously healthy children. METHODS: To determine the prevalences of 11 viruses--respiratory syncytial virus (RSV), adenovirus, rhinovirus, parainfluenza viruses (PIVs) 1 and 3, influenza viruses A and B, hMPV, HCoV, HCoV-NL63, and HBoV--among infants or children with LRTIs, in association with their epidemiologic characteristics, we performed multiplex reverse-transcriptase polymerase chain reaction on nasopharyngeal aspirates obtained from 515 children < or =5 years old with LRTIs during the period 2000-2005. RESULTS: Viruses were identified in 312 (60.6%) of the 515 patients. RSV was detected in 122 (23.7%), HBoV in 58 (11.3%), adenovirus in 35 (6.8%), PIV-3 in 32 (6.2%), rhinovirus in 30 (5.8%), hMPV in 24 (4.7%), influenza A in 24 (4.7%), PIV-1 in 9 (1.7%), influenza B in 9 (1.7%), and HCoV-NL63 in 8 (1.6%). Coinfections with > or =2 viruses were observed in 36 patients (11.5%). Twenty-two patients (37.9%) infected with HBoV had a coinfection. Bronchiolitis was frequently diagnosed in patients who tested positive for RSV, PIV-3, or rhinovirus, whereas influenza A, PIV-1, and HCoV-NL63 were commonly found in patients with croup. The age distributions of patients with viral infections differed; notably, RSV was responsible for 77% of LRTIs that occurred in infants < or =3 months old. The number of hMPV infections peaked between February and April, whereas the number of HCoV-NL63 infections peaked between April and May. CONCLUSIONS: This study describes the features of LRTIs associated with newly identified viruses in children, compared with those associated with known viruses. Additional investigations are required to define the role of HBoV in LRTI.

Child, Preschool↗

Etiologic and clinical evaluation of acute lower respiratory tract infections in young Argentinian children: an overview.

This paper summarizes the first study on clinical, etiologic, and epidemiologic features of acute lower respiratory tract infection (ALRI) in children in Argentina. A total of 1,003 children less than 5 years of age (805 inpatients and 198 outpatients) presenting with ALRI were studied during a 40-month period. Nasopharyngeal aspirate (NPA), blood, urine, and throat-swab samples were collected when each child was first seen for care. Virologic studies were performed on the NPA by means of indirect immunofluorescence and isolation of virus in cell culture. Bacteriologic studies primarily were done by means of culture of blood or pleural fluid (when available); Bordetella pertussis and Mycoplasma pneumoniae, however, were searched for by the use of immunofluorescence and complement-fixation testing, respectively, in paired sera. Respiratory syncytial virus was the most commonly isolated virus, followed by adenovirus, parainfluenza virus, and influenza virus. Streptococcus pneumoniae was the most frequently isolated bacterium, followed by B. pertussis and Haemophilus influenzae type b. Overall, the patient fatality rate was 3.8% among inpatients with pneumonia or bronchiolitis.

Acute Disease↗

Identification of viruses in a study of acute respiratory tract infection in children from Uruguay.

For the purpose of identifying viral agents associated with acute respiratory tract infections (ARI) in children less than 5 years old, a longitudinal community study was undertaken in Montevideo, Uruguay, from May 1985 to December 1987. This report includes results obtained by cell culture and immunofluorescence techniques for detection of respiratory syncytial virus (RSV), influenza A and B viruses, parainfluenza 1 and 3 viruses, and adenovirus. Two populations were studied: children visited at home by pediatricians (group 1) and children with an ARI episode who attended an outpatient clinic (group 2). Nasopharyngeal aspirates were obtained at the time of an ARI episode: 858 from group 1 and 488 from group 2. Viruses were identified in 15.3% of group 1 specimens and in 17.6% of group 2 specimens. RSV was the most frequently recovered agent, accounting for 67.9% and 58.1%, respectively, of all viruses detected. The sensitivity and specificity of RSV isolation by cell culture are compared with detection by indirect immunofluorescence.

Acute Disease↗

Community respiratory virus infections among hospitalized adult bone marrow transplant recipients.

From 1 November 1992 through 1 May 1993 and from 1 November 1993 through 1 May 1994, we conducted a prospective surveillance study at the University of Texas M.D. Anderson Cancer Center (Houston) to evaluate the role of community respiratory virus infections in hospitalized adult bone marrow transplant (BMT) recipients, Respiratory secretions were obtained from all adult BMT recipients with acute respiratory illnesses. During these two winters, a community respiratory virus was isolated from 37 (36%) of 102 patients and 30 (26%) of 115 patients, respectively. Approximately half (49%) of these infections were due to respiratory syncytial virus (RSV); the remainder were due to influenza virus (18%), picornaviruses (18%), parainfluenza virus (9%), or adenovirus (6%). Fifty-eight percent of these infections were complicated by pneumonia, with an associated mortality of 51%. The pneumonias that complicated RSV infection were almost exclusively viral in origin and were associated with a mortality of 100% if not treated promptly with antiviral agents. In contrast, many of the pneumonias that complicated the other viral infections, such as influenza, appeared to be either self-limited viral pneumonias or secondary bacterial or fungal pneumonias. Community respiratory viruses are frequent causes of acute respiratory illnesses in adult BMT recipients hospitalized during the winter and are associated with substantial morbidity and mortality.

Adenovirus Infections, Human↗

Epstein-Barr virus infections in Brazil. II. Hodgkin's disease.

Sixty-seven cases of Hodgkin's disease (HD) occurring in São Paulo, Brazil, were studied. Males with HD predominated over females 2.3 to 1. Sixty-six percent of the cases occurred in patients under 30 years of age, 31.7% under 20 years of age, and only 7.5% after 50 years of age. Lymphocyte predominance and mixed cellularity histologic types were most common in patients less than 15 years old, and nodular sclerosis was most common in the 15- to 19-year-old group. Sera from all patients had antibody to the viral capsid antigen (VCA) of Epstein-Barr virus (EBV). The geometric mean titer (GMT) of VCA antibody with the use of Jijoye cells as antigen was 1:162, and 31.3% of patients had titers of 1:320 or more; in controls, the GMT was 1:67 and 3.8% had titers of 1:320 or more. Similar results were obtained when EB-3 cells were used as antigen. The highest titers occurred in males, in mixed cellularity and lymphocyte depletion forms, and in stage 2 of illness. EBV-specific IgM antibody and heterophile antibody levels were not elevated, but 20.5% of the HD patients had antibody to the early antigen of EBV present in their sera. Antibody levels for herpes simplex virus, cytomegalovirus, rubella, measles, parainfluenza viruses, and papovavirus were not significantly elevated over those in matched controls.

Adolescent↗

The Tucson Children's Respiratory Study. II. Lower respiratory tract illness in the first year of life.

Lower respiratory tract illnesses occurring during the first year of life in 1,179 healthy infants enrolled in the Children's Respiratory Study, Tucson, Arizona, are described. The children, who use the pediatricians of a health maintenance organization, were enrolled into the study between May 1980 and January 1984. Data were collected on signs, symptoms, and diagnosis for each illness; nasopharyngeal and throat swabs were collected at the acute visit for viral, chlamydial, and mycoplasmal cultures. The cumulative incidence of illness in the first year of life was 32.88 per 100 children. Of the 348 initial lower respiratory tract illnesses occurring in these infants, 60% were diagnosed as bronchiolitis. At least one infecting agent was identified in 66% of the specimens collected at the time of the first illness. Respiratory syncytial virus was the most common isolate; 12 other agents were also identified. There was a strong (p less than 0.0001) relation between agent identified, symptoms reported, and diagnosis; bronchiolitis was predominantly associated with respiratory syncytial virus and croup with parainfluenza viruses. Sex and ethnicity were unrelated to illness experience or to characteristics of the first illness. Lower respiratory tract illness occurrence in the Children's Respiratory Study appears to be similar to patterns observed elsewhere, suggesting that diagnoses (and infecting agents) have changed little over the past decades.

Humans↗