Sample type is crucial to the diagnosis of Mycoplasma pneumoniae pneumonia by PCR.

Sensitive and specific methods for rapid laboratory diagnosis of Mycoplasma pneumoniae were not available until nucleic acid amplification methods were developed. The choice of sample type and method of sampling are crucial to optimal diagnostic efficacy. Three types of respiratory samples from 32 young military conscripts with pneumonia were collected during an outbreak of M. pneumoniae infection. Sputum, nasopharyngeal aspirate and throat swab specimens were tested by 16S rRNA gene-based PCR with liquid-phase probe hybridization, and the results were compared with serology. The PCR result was positive for 22 (69 %) of the sputa, 16 (50 %) of the aspirates and 12 (37.5 %) of the swabs. Serology with increasing or high titres supported the positive findings in all instances. Sputum, when available, is clearly the best sample type for young adults with pneumonia.

Viral etiology of frequently recurring respiratory tract infections in children.

The viral etiology of frequently recurring respiratory tract infection (FRRI) in children aged <2 years was studied. Altogether, 329 children were followed from 2 to 24 months of age in the Finnish Otitis Media Cohort Study. Children with FRRI were defined as having > or =9 episodes of upper respiratory tract infection (URI) and/or > or =4 episodes of acute otitis media during follow-up. Nasopharyngeal aspirates, middle ear fluid specimens, and serum samples were analyzed for 8 common respiratory viruses. Of 1358 URI episodes, 642 (47%) occurred in the 78 children with FRRI. At least 1 virus was associated with 62% of these episodes, whereas the corresponding figure for children without FRRIs was 54%. The frequency of different viruses was similar in both groups, but the relative proportion of rhinovirus infections was slightly higher among children with FRRI. In conclusion, a specific viral etiology does not explain the excess of URI episodes in children with FRRI.

No evidence of Mycoplasma pneumoniae in acute myringitis.

Our aim was to discover Mycoplasma pneumoniae in bullous and hemorrhagic myringitis in children <2 years of age. Middle ear fluid samples (n = 37) and samples taken from the blisters of the tympanic membranes (n = 12) studied by polymerase chain reaction for M. pneumoniae were negative. This study does not support an important role for M. pneumoniae as an etiologic agent in acute myringitis.

Evaluation of the role of respiratory viruses in acute myringitis in children less than two years of age.

BACKGROUND: The etiology of acute myringitis remains controversial although it is usually encountered in connection with acute otitis media (AOM). In most cases of acute myringitis a bacterial pathogen has been detected in the middle ear fluid, but the role of respiratory viruses has remained unclear. Our objective was to investigate the etiologic role of viruses in the pathogenesis of acute bullous and hemorrhagic myringitis in children <2 years of age. METHODS: A prospective longitudinal cohort study of 2028 children ages 7 to 24 months in primary care in the Finnish Otitis Media Vaccine Trial. Nasopharyngeal aspirate (NPA) and middle ear fluid (MEF) samples taken at the time of the diagnosis were examined by a time-resolved fluoroimmunoassay for antigen detection of adenoviruses; influenza viruses A and B; parainfluenza viruses 1, 2 and 3; and respiratory syncytial virus and by reverse transcription polymerase chain reaction for human rhinovirus and human enterovirus. RESULTS: Eighty-two children with bullous myringitis and 37 children with hemorrhagic myringitis were diagnosed during the 18-month follow-up period. In bullous myringitis a respiratory virus was detected in 70% of NPA samples and in 27% of MEF samples. In hemorrhagic myringitis 57% of NPA samples and 28% of MEF samples were virus-positive. The viral distribution was similar to that of AOM (virus positive 64% of NPA and 37% of MEF). CONCLUSIONS: We could not confirm any specific respiratory virus to be the etiologic agent in acute myringitis. The etiology of acute myringitis is similar to that of AOM in children <2 years of age.

The value of virus serology in epidemiological studies of acute otitis media in children.

BACKGROUND: Acute otitis media (AOM) is a major health problem in young children. There is a general conception that AOM is a bacterial disease but with the availability of sensitive diagnostic methods, it has gradually become evident that viruses play an important role in the pathogenesis of AOM. Paired blood samples are seldom taken from infants although valuable information could be obtained by serological methods. During the recent Finnish Otitis Media (FinOM) Cohort Study, in addition to nasopharyngeal aspirates (NPA) and middle ear fluids (MEF), paired acute and convalescent serum samples were collected from children with AOM. OBJECTIVES: To establish the diagnostic value of serological methods in etiological and epidemiological studies of AOM. STUDY DESIGN: A complete set of NPA, MEF, and paired sera was collected during 447 events of AOM experienced by 179 children between 2 months and 2 years of age. Antigens of respiratory syncytial virus (RSV), adenoviruses, influenza A and B, and parainfluenza types 1-3 in NPAs and MEFs were detected by time-resolved fluoroimmunoassay (TR-FIA), and antibody titers were determined by complement fixation test (CFT) or by enzyme immunoassay. RESULTS: A total of 163 virus-positive events were identified. Of those, only 34 were positive by TR-FIA and by serology. From 48 events a positive result was obtained only by TR-FIA and from 81 only by serology. CONCLUSION: Although serological methods are usually of little use in clinical practice, epidemiological studies clearly gain value if serology is included. The number of virus-positive findings dramatically increased by including serological tests in the diagnostic work-up of these AOM events.

Is there any specific association between respiratory viruses and bacteria in acute otitis media of young children?

BACKGROUND: Respiratory viral infections are usually preceding or coinciding with acute otitis media (AOM) in children. It is not known if a given viral infection would facilitate invasion of bacterial pathogens into the middle ear in a species-specific way. We reanalysed the microbiological results of the two prospective Finnish Otitis Media (FinOM) studies for this purpose. METHODS: The children had been followed from 2 months to 2 years of age in specific study clinics and all referred AOM events were analysed. Combined results of virus detection tests from middle ear fluid and nasopharyngeal aspirate and those of bacterial culture from middle ear fluid were cross-tabulated for 529 AOM events in the FinOM Cohort Study and for 364 events in the FinOM Vaccine Trial. RESULTS: In both studies the main bacterial pathogens were Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis while the main viruses detected were rhinoviruses and respiratory syncytial virus (plus enteroviruses in the Vaccine Trial). No distinct species-specific associations were observed between the viral and bacterial findings. CONCLUSION: We did not find support to the theory that respiratory infection caused by a given viral species would favour growth of a certain bacterial pathogen in the MEF more than another.

Aetiology of community-acquired pneumonia: serological results of a paediatric survey.

Serological methods are routinely used in the diagnosis of viral and atypical bacterial respiratory infections. Recently, they have also been applied to typical bacteria, such as Streptococcus pneumoniae. The aim of this study was to determine the aetiology of paediatric community-acquired pneumonia (CAP) in both ambulatory and hospitalized patients, by using antibody assays. During a 15-month prospective surveillance, paired sera were studied for antibodies to 14 microbes in 101 children with symptoms of acute infection and infiltrates compatible with pneumonia on chest radiographs. A potential causative agent was detected in 66 (65%) patients. Evidence of bacterial, viral and mixed viral-bacterial infection was demonstrated in 44%, 42% and 20% of the CAP cases, respectively. The most commonly found agents included Mycoplasma pneumoniae (27%), Pneumococcus (18%) and respiratory syncytial virus (17%). Human metapneumovirus (hMPV) was detected in 5 (5%) children. Pneumococcal infections were evenly distributed among the age groups studied. Our results confirm the role of S. pneumoniae in paediatric CAP at all ages, those of M. pneumoniae at >2 y of age and emphasize the emerging role of hMPV. The high proportion of mixed viral-bacterial infections highlights the need to treat all children with CAP with antibiotics.

Incidence of community-acquired pneumonia in children caused by Mycoplasma pneumoniae: serological results of a prospective, population-based study in primary health care.

OBJECTIVE: The objective of the present study was to assess the incidence of community-acquired pneumonia (CAP) in children caused by Mycoplasma pneumoniae. METHODOLOGY: During 12 months in 1981-1982, all CAP cases in a defined child population were registered. M. pneumoniae aetiology, initially measured by complement fixation (CF) test, was in 1999 supplemented by measurement of IgG and IgM antibodies using enzyme immunoassays (EIA). RESULTS: M. pneumoniae was detected in 61 (30%) of 201 paediatric CAP cases, being the most common aetiological agent in those 5 years of age or over. At that age, M. pneumoniae was responsible for over 50% of cases, and over 90% of mycoplasmal cases were treated as outpatients. The EIA detected 17 new cases over and above the 44 detected by CF, while CF alone revealed 10 cases. The incidence of M. pneumoniae CAP increased with age, being over 10/1000 children at the age of 10 years or more. Co-infections with Streptococcus pneumoniae and Chlamydia pneumoniae were present in over 30% and 15%, respectively, of mycoplasmal CAP cases. CONCLUSION: M. pneumoniae is a common cause of paediatric CAP in primary health care, and co-infections with S. pneumoniae are common. Both S. pneumoniae and M. pneumoniae should be taken into account when starting antibiotics for children with CAP.

Influence of antigenic drift on the intensity of influenza outbreaks: upper respiratory tract infections of military conscripts in Finland.

A total of 102,600 upper respiratory infections (URI) were recorded among young military conscripts in the Finnish Defence Forces during the study period from October 1991 to March 1994. This period covered three outbreaks caused by H3N2-subtype influenza A virus and one outbreak of influenza B. During the 1991/92 outbreak caused by A/Beijing/353/89-like virus, the calculated influenza A incidence was 2,206/10,000 men. During the 1992/93 outbreak when influenza B was the predominant virus, a new drift variant of influenza A that belonged to the lineage of A/Beijing/32/92-like and A/Shangdong/9/93-like viruses circulated but the incidence of influenza A was not more than 1,044/10,000. A higher incidence, 2,810/10,000, was recorded during the 1993/94 outbreak, when the circulating virus was similar to the 1992/93 virus antigenically and with regard to haemagglutinin and neuraminidase (NA) gene sequences. Crossreactive haemagglutination-inhibition antibodies induced in 1991/92 probably were sufficient to restrict the epidemic activity in 1992/93 but no longer in 1993/94. Furthermore, during the 1991/92 outbreak, some of the A/Beijing/353/89-like viruses already had shared the NA sequence markers characteristic of the viruses in 1992/93 and 1993/94, which may also have strengthened protection in 1992/93.

Presence of specific viruses in the middle ear fluids and respiratory secretions of young children with acute otitis media.

The purpose of the study was to investigate the presence of different viruses in middle ear fluids and nasopharyngeal aspirates in young children with acute otitis media. Two cohorts of children (N = 329 and 611) were followed from 2 to 24 months of age in Finland in two prospective studies (Finnish Otitis Media Cohort Study and Finnish Otitis Media Vaccine Trial). During the study period, nasopharyngeal and middle ear fluid specimens for each acute otitis media event were examined for eight (Cohort Study) or ten (Vaccine Trial) common respiratory viruses; adenoviruses, influenza viruses A and B, parainfluenza viruses 1, 2, and 3, respiratory syncytial virus (RSV), enteroviruses, parechoviruses, and rhinoviruses. Picornaviruses (rhinoviruses, enteroviruses, and parechoviruses) were determined by reverse transcription PCR while antigen detection was used for the other viruses. A virus was present in either nasopharyngeal or middle ear specimen in 54% of events in the first cohort and in 67% of events in the second. Rhinoviruses formed the most common virus group detected (41-32%), followed by enteroviruses (25%, sought in the second cohort only) and respiratory syncytial virus (RSV) (10%). All the other viruses represented jointly 8-10% of the events. In conclusion, using the methods described in this study, a specific virus infection was diagnosed in two thirds of all acute otitis media events in young children. Picornavirus RNA was detected in association with more than a half of all acute otitis media events. The use of PCR-based methods for the other respiratory viruses might have increased further the overall virus detection rate in acute otitis media.