Search PubMed⌕ Search

Biomedical subjects

M Leinonen

Publications and source records attributed to M Leinonen.

At least 145 records · Page 8Linked to original sources

Comparison of PCR assay with bacterial culture for detecting Streptococcus pneumoniae in middle ear fluid of children with acute otitis media.

We have studied etiological diagnosis of acute otitis media (AOM) by comparing a newly developed pneumococcal PCR for Streptococcus pneumoniae to bacterial culture with 180 middle ear fluid (MEF) samples of 125 children with 125 episodes of AOM. For pneumococcal PCR assay, DNA from MEF samples was extracted by phenol-chloroform. The outer primers used amplified a 348-bp region of the pneumolysin gene, and the inner primers amplified a 208-bp region. S. pneumoniae was cultured in 33 (18%) samples, and pneumolysin PCR was positive for 51 (28%) of 180 MEF samples. Only 2 of 21 PCR-positive, S. pneumoniae culture-negative samples were positive for other otitis pathogens. By combining MEF culture and PCR results, 54 (30%) of 180 MEF samples had evidence of pneumococcal etiology. In conclusion, pneumolysin PCR is a sensitive and specific new method to study pneumococcal involvement in MEF samples of children with AOM.

Acute Disease↗

Pneumococci in nasopharyngeal samples from Filipino children with acute respiratory infections.

The presence of Streptococcus pneumoniae in the upper respiratory tract was studied in 318 Filipino children less than 5 years old with an acute lower respiratory tract infection. Nasopharyngeal samples were obtained from 292 children. With both quantitative bacterial culture and detection of capsular polysaccharide antigens by coagglutination, counterimmunoelectrophoresis, and latex agglutination, pneumococci were found in 160 (70%) of the 227 samples eligible for analysis. Culture was positive in 115 samples and antigen was positive in 140 samples. The culture isolation rate was significantly lower if the patient had received antimicrobial agents in the 48 h prior to the sampling. The seven most common types or groups of pneumococci were 6, 14, 19, 23, 15, 7, and 11, which together accounted for 64% of all pneumococcal findings.

Acute Disease↗

Interference of immunoglobulin G (IgG) antibodies in IgA antibody determinations of Chlamydia pneumoniae by microimmunofluorescence test.

In the microimmunofluorescence test for measuring immunoglobulin A (IgA) antibodies against Chlamydia pneumoniae, removal of interfering IgG antibodies made IgA antibody reactivity patterns in 952 serum samples easier to interpret, prozone effects disappeared, and titers increased, especially in the sera with high IgG titers. IgA rheumatoid factors did not interfere in the assay.

Adult↗

Moraxella catarrhalis--an uncommon cause of community-acquired pneumonia in Swedish children.

In 284 Swedish children with community-acquired, roentgenologically verified pneumonia, antibodies to Moraxella (Branhamella) catarrhalis were determined in paired serum samples with an enzyme immunoassay using a whole-cell antigen preparation from 10 strains of M. catarrhalis. Only 9 children (3%) had significant increases in antibodies to M. catarrhalis. Among these 9 children, 11-39 months of age, 6 had serologic evidence of concurrent infection with other respiratory pathogens such as S. pneumoniae, non-capsulated H. influenza, RS virus and adenovirus. In 6 (67%) of the 9 children with antibody response and in 74 (27%) of the 275 children without antibody response to M. catarrhalis, nasopharyngeal cultures yielded growth of this bacterium. M. catarrhalis seems to be a common commensal in the upper respiratory tract, but a rare cause of pneumonia in children.

Antibodies, Bacterial↗

Serological diagnosis of pneumococcal pneumonia--will it ever become a clinical reality.

Bacterial serology is not commonly used in the etiological diagnosis of pneumococcal pneumonia. This is partly because of the lack of commercial tests for this purpose. In addition, if paired sera taken about 2 weeks apart are needed, the positive pneumococcal finding obtained after the patient has been discharged from the hospital does not help clinicians in selecting the proper antibiotic treatment for the patient. Pneumococcal serology, however, is needed in both etiologic and epidemiological surveys as well as in the studies on the clinical efficacy of pneumococcal vaccines.

Antibodies, Bacterial↗

Experimental infection of Chlamydia pneumoniae in mice.

NIH/S, Swiss Webster, and BALB/c mice were infected intranasally with three Chlamydia pneumoniae isolates, Kajaani 6, Helsinki 12, and TW-183. C. pneumoniae could be isolated from the lung homogenates and bronchoalveolar lavage fluids up to the third week post-infection. Specific serum IgG antibodies against C. pneumoniae reached high levels in the third week and remained elevated until the end of the 6-week follow-up period. Serum IgM levels were highest in the third week post-infection and started to decrease thereafter. In spite of these signs of ongoing infection, the mice did not show any symptoms of disease. NIH/S mice could be readily and uniformly infected, while BALB/c mice were the most resistant and developed the weakest antibody response. The greatest histological changes were detected in NIH/S mice as well. The inflammatory infiltrate, which consisted of lymphocytes and plasma cells throughout the study, was restricted to the peribronchial and perivascular space and to the interstitium of the lung parenchyma.

Animals↗

Aetiology of community-acquired pneumonia in children treated in hospital.

Viral and bacterial antigen and antibody assays were prospectively applied to study the microbial aetiology of community-acquired pneumonia in 195 hospitalised children during a surveillance period of 12 months. A viral infection alone was indicated in 37 (19%), a bacterial infection alone in 30 (15%) and a mixed viral-bacterial infection in 32 (16%) patients. Thus, 46% of the 69 patients with viral infection and 52% of the 62 patients with bacterial infection had a mixed viral and bacterial aetiology. Respiratory syncytial virus (RSV) was identified in 52 patients and Streptococcus pneumoniae in 41 patients. The next common agents in order were non-classified Haemophilus influenzae (17 cases), adenoviruses (10 cases) and Chlamydia species (8 cases). The diagnosis of an RSV infection was based on detecting viral antigen in nasopharyngeal secretions in 79% of the cases. Pneumococcal infections were in most cases identified by antibody assays; in 39% they were indicated by demonstrating pneumococcal antigen in acute phase serum. An alveolar infiltrate was present in 53 (27%) and an interstitial infiltrate in 108 (55%) of the 195 patients. The remaining 34 patients had probable pneumonia. C-reactive protein (CRP), erythrocyte sedimentation rate and total white blood cell count were elevated in 25%, 40% and 36% of the patients, respectively. CRP was more often elevated in patients with bacterial infection alone than in those with viral or mixed viral-bacterial infections. No other correlation was seen between the radiological or laboratory findings and serologically identified viral, bacterial or mixed viral-bacterial infections.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Evaluation of serological methods in the diagnosis of Chlamydia pneumoniae pneumonia during an epidemic in Finland.

A complement fixation (CF) test, a micro-immunofluorescence (micro-IF) test and an enzyme immunoassay (EIA) using Re-lipopolysaccharide as antigen were compared in the diagnosis of chlamydial infection in 136 mainly elderly patients hospitalized with community-acquired pneumonia during a Chlamydia pneumoniae epidemic in Finland in 1986-1987. Chlamydial pneumonia was diagnosed in 58 (42.6%) of the 136 pneumonia patients; 44 (75.9%) of them could be shown by micro-IF to be caused by Chlamydia pneumoniae, three by Chlamydia psittaci and four by Chlamydia spp. Only 5 (11.4%) of 44 patients with Chlamydia pneumoniae pneumonia were IgM-positive, indicating that the majority of cases were reinfections. In this population of mainly elderly patients the CF test was insensitive, being positive in only 6 (10.3%) of 58 cases of chlamydial pneumonia. The EIA detected 72.4% of cases and micro-IF 87.9% of cases (including infections with Chlamydia pneumoniae, Chlamydia psittaci and Chlamydia spp.). In the EIA 77% of positive cases were positive in serum samples taken a week apart, whereas the corresponding figure for micro-IF was 50%. In micro-IF the measurement of IgA antibody levels is recommended and IgM-positive sera should be retested after removal of IgG antibody to avoid false-positive findings due to presence of rheumatoid factor. The collection of a third serum sample, for instance one month after onset, is also recommended, since half of the patients showed a diagnostic response in the micro-IF only in the sera taken one month apart.

Adolescent↗

Use of nasopharyngeal isolates of Streptococcus pneumoniae and Haemophilus influenzae from children in Pakistan for surveillance for antimicrobial resistance.

Antimicrobial resistance of Streptococcus pneumoniae and Haemophilus influenzae presents a challenge to clinical case management, particularly in programs for acute respiratory tract infection (ARI), including pneumonia, in developing countries. To determine whether nasopharyngeal isolates of S. pneumoniae and H. influenzae from a clinically defined group of children could be used to predict the prevalence of antimicrobial resistance of strains that cause disease, 601 urban children with ARI, 133 healthy urban children and 285 rural children were evaluated in Pakistan. Of the urban children with ARI, 216 (35.9%) were bacteremic, predominantly with S. pneumoniae (108 children) and H. influenzae (100 children). Overall 631 (61.9%) children carried S. pneumoniae and 381 (37.4%) carried H. influenzae. The proportions of nasopharyngeal isolates of both organisms from urban children with ARI resistant to penicillin or ampicillin, trimethoprim/sulfamethoxazole, chloramphenicol and erythromycin were similar to the proportions of resistant blood isolates. Nasopharyngeal isolates from rural children had lower rates of resistance to some antimicrobial agents. These findings suggest that nasopharyngeal isolates of S. pneumoniae and H. influenzae from children with ARI can be used to conduct surveillance for antimicrobial resistance in a defined geographic area. Such surveillance would aid programs in developing countries in making a rational choice of antimicrobial agents for use in clinical management of bacterial diseases, including pneumonia.

Acute Disease↗

Antigen and antibody assays in the aetiological diagnosis of respiratory infection in children.

The diagnostic efficacy of two methods--demonstration of seroconversion in paired sera and detection of antigen in clinical specimens--was evaluated in 183 children with respiratory syncytial, parainfluenza or adenoviral, or pneumococcal respiratory tract infection. Viral infection was diagnosed in 46 (37%) of the 125 cases by antigen assay alone, in 36 (29%) by antibody assay alone and in 43 (34%) by both methods. In respiratory syncytial viral infections, antigen assays were more often positive than antibody assays; 80% of the cases were antigen positive and 63% solely antigen positive. In parainfluenza and adenoviral infections, antigen assays were not as useful; a positive result was seen in 59% and 44% of cases, respectively. Pneumococcal infection was diagnosed in 25 (30%) of the 84 cases by antigen assay alone, in 54 (64%) by antibody assay alone and in only 5 (6%) by both methods. Thus nearly all pneumococcal infections were diagnosed by only one method. The efficacy of antigen and antibody assay was clearly dependent on the age of the patients. In infants less than six months of age, nearly all (27 (90%)) of the 30 infections were diagnosed by antigen detection; 25 solely by antigen detection. In older patients, antigen and antibody assays supplemented each other. We conclude that antigen detection should be used as the primary method for the diagnosis of viral or pneumococcal respiratory tract infection. In infants, antigen detection is the only reliable method of microbial diagnosis. In addition, antigen detection is the method of rapid microbial diagnosis. In pneumococcal infections, the sensitivities of antigen detection methods are not sufficient; this is an important area for further research.

Adolescent↗

Fever and neutropenia: bacterial etiology revealed by serological methods.

In a prospective study, 91 episodes of fever in neutropenic children with cancer were evaluated. Fifteen episodes were septicemias, verified by a positive blood culture, 62 were fevers of unknown origin, 6 were focal infections and 8 were of other etiologies (i.e. drug fevers and viral infections). Serum antibody responses to bacteria were measured in paired sera by an enzyme immunoassay method. Bacterial infection was demonstrated serologically in 20% of documented septicemias, in 35% of fevers of unknown origin and occasionally in the other groups. Tests were available and found positive in the fever of unknown origin group for Staphylococcus aureus, Streptococcus pneumoniae, Haemophilus influenzae, Moraxella (Branhamella) catarrhalis and enterobacteria. Some had multiple etiology. In conclusion, bacterial serology is a promising method of identifying bacterial etiology in fever of otherwise unknown origin in neutropenic children with cancer.

Adolescent↗

Cell-mediated immunity to Chlamydia pneumoniae measured as lymphocyte blast transformation in vitro.

The purpose of the present study was to analyze Chlamydia pneumoniae-induced, antigen-specific, cell-mediated immunity. Peripheral blood mononuclear cells of four persons infected with C. pneumoniae Kajaani 6 and 17 healthy volunteers were stimulated with antigen composed of whole elementary bodies of C. pneumoniae Kajaani 6 (CP-Ag). Definitive antigen-specific lymphoproliferation (LP) responses were developed after recent infection. The LP responses of healthy people to CP-Ag varied considerably. There was no clear correlation between LP responses to CP-Ag and those to an antigen prepared from Chlamydia trachomatis serotype L2 (r > or = 0.50, P < 0.1). A larger study is required to demonstrate whether the LP responses to CP-Ag can be used for the diagnosis of C. pneumoniae infection.

Adult↗

Mapping of immunoreactive sites of pneumococcal pneumolysin by use of synthetic peptides.

In the search for better diagnostic tools and improved vaccines against pneumococcal diseases, continuous antigenic sites of the pneumococcal pneumolysin molecule were identified with partially overlapping synthetic peptides. Peptides were derived from the predicted amino acid sequence according to the known DNA sequence of pneumolysin. The immunoreactivities of the peptides were studied by enzyme immunoassay with seven human serum samples and three rabbit hyperimmune serum samples. Two immunoreactive sites in the overlapping peptides at the amino-terminal end of the pneumolysin molecule were identified in this study. Two common immunoreactive sites for both human and rabbit sera were demonstrated.

Amino Acid Sequence↗

New method to assess dilution of secretions for immunological and microbiological assays.

Accurate quantitation of pathogens and antibody concentrations in secretions has been difficult because of unpredictable dilution of secretion with th diluent at the time of sample collection. We added an inert substance, lithium chloride (LiCl), to the sample diluent and measured its concentration with an atomic absorption spectrometer before and after the specimen was added. LiCl, at a concentration of 2 mmol of Li per liter, has no negative effect on the survival of common respiratory pathogens or on the results of immunoassays. The method is applicable to any sample collecting in which dilution of the specimen is necessary.

Antibodies↗

Chlamydia pneumoniae-specific circulating immune complexes in patients with chronic coronary heart disease.

BACKGROUND: An association of chronic Chlamydia pneumoniae infection to coronary heart disease has been suggested recently. In a recent study, we demonstrated circulating immune complexes containing chlamydial genus-specific lipopolysaccharide in patients with coronary heart disease. The objective of the present study was to investigate whether C. pneumoniae species-specific immune complexes are present in chronic coronary heart disease. METHODS AND RESULTS: The presence of Chlamydia-specific circulating immune complexes was studied in 46 patients with chronic coronary heart disease and in control subjects. Chlamydial lipopolysaccharide-containing immune complexes were detected with the antigen-specific capture method, and they were present in 41% of patients and 15% of control subjects (p < 0.01). The presence of C. pneumoniae antibodies in circulating immune complexes was studied by testing the specificity of antibodies derived from isolated and dissociated immune complexes by microimmunofluorescence testing and immunoblotting. The C. pneumoniae indexes based on the relative amount of immune complex-derived antibodies and free antibodies were significantly higher among patients compared with control subjects (median, 1/8 versus 1/16; p < 0.001). Immune complex bound antibodies showed specificity for 98-kd and 42-kd proteins of C. pneumoniae. CONCLUSIONS: The results suggest that the majority of the patients with chronic coronary heart disease have a chronic C. pneumoniae infection in which chlamydial components have an easy access to circulation to form immune complexes with preexisting antibodies. These findings give further evidence for the association of chronic C. pneumoniae infection with coronary heart disease.

Antigen-Antibody Complex↗

Mycoplasma pneumoniae and Chlamydia trachomatis in acute lower respiratory infections in Filipino children.

The role of Mycoplasma pneumoniae and Chlamydia trachomatis in acute lower respiratory infection (ALRI) was studied by serologic analysis of 216 Filipino children less than five years old living in periurban slums and middle-class neighborhoods. Diagnostic antibody responses to M. pneumoniae and C. trachomatis were observed in 18 (8.3%) and five (4.3%), respectively, of the cases with ALRI. These bacteria are not susceptible to the antimicrobial treatments currently recommended for the treatment of ALRI in developing countries.

Acute Disease↗