ABSTRACT
An enzyme-linked immunosorbent assay specific to outer membrane protein P6 (P6-ELISA) was applied for detecting Haemophilus influenzae in middle ear fluids (MEFs) from acute otitis media (AOM) patients and in nasopharyngeal secretions (NPSs) from acute rhinosinusitis patients. P6-ELISA had a sensitivity of 83.3% for MEFs and 71.5% for NPSs and a specificity of 85.6% for MEFs and 92.5% for NPSs, respectively. Real-time PCR exhibited significant differences in the number of ompP1 gene copies among samples determined by P6-ELISA to be positive and negative for H. influenzae. However, because the P6-ELISA test has the reactivity in Haemophilus species include two commensals H. haemolyticus and H. parainfluenzae, it is thus a weak method in order to detect only NTHi correctly. Consequently, diagnosis using the P6-ELISA should be based on an overall evaluation, including the results of other related examinations and clinical symptoms to prevent misleading conclusions in clinical setting.
Subject(s)
Bacterial Outer Membrane Proteins/metabolism , Haemophilus Infections/diagnosis , Haemophilus Vaccines/metabolism , Haemophilus influenzae/metabolism , Otitis Media/diagnosis , Rhinitis/diagnosis , Sinusitis/diagnosis , Acute Disease , Adolescent , Adult , Aged , Bacterial Outer Membrane Proteins/genetics , Child , Child, Preschool , Ear, Middle/microbiology , Enzyme-Linked Immunosorbent Assay , Female , Haemophilus Infections/microbiology , Haemophilus Vaccines/genetics , Haemophilus influenzae/genetics , Humans , Infant , Male , Middle Aged , Nasopharynx/microbiology , Otitis Media/microbiology , Rhinitis/microbiology , Sensitivity and Specificity , Sinusitis/microbiology , Young AdultABSTRACT
Since the incidence of penicillin-resistant Streptococcus pneumoniae has been increasing at an astonishing rate throughout the world, the need for accurate and rapid identification of pneumococci has become increasingly important to determine the appropriate antimicrobial treatment. We have evaluated an immunochromatographic test (ODK-0901) that detects pneumococcal antigens using 264 middle ear fluids (MEFs) and 268 nasopharyngeal secretions (NPSs). A sample was defined to contain S. pneumoniae when optochin and bile sensitive alpha hemolytic streptococcal colonies were isolated by culture. The sensitivity and specificity of the ODK-0901 test were 81.4% and 80.5%, respectively, for MEFs from patients with acute otitis media (AOM). In addition, the sensitivity and specificity were 75.2% and 88.8%, respectively, for NPSs from patients with acute rhinosinusitis. The ODK-0901 test may provide a rapid and highly sensitive evaluation of the presence of S. pneumoniae and thus may be a promising method of identifying pneumococci in MEFs and NPSs.
Subject(s)
Chromatography, Affinity , Nasopharynx/metabolism , Otitis Media with Effusion/diagnosis , Pneumococcal Infections/diagnosis , Rhinitis/diagnosis , Sinusitis/diagnosis , Streptococcus pneumoniae/isolation & purification , Teichoic Acids/metabolism , Acute Disease , Adolescent , Adult , Bacterial Proteins/genetics , Case-Control Studies , Child , Child, Preschool , Ear, Middle/microbiology , Female , Humans , Infant , Infant, Newborn , Male , Middle Aged , Otitis Media with Effusion/microbiology , Pneumococcal Infections/microbiology , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Rhinitis/microbiology , Sensitivity and Specificity , Sinusitis/microbiology , Streptococcus pneumoniae/immunology , Teichoic Acids/immunology , Young AdultABSTRACT
To determine the distribution of Streptococcus pneumoniae serotypes isolated from patients under 6 years of age with acute suppurative otitis media, to calculate the serotype coverage of 7-valent pneumococcal conjugate vaccine, and to clarify trends in PCG-resistant Streptococcus pneumoniae, we conducted a one-year prospective study from April 2005 to March 2006 at 10 medical institutions in Hokkaido, Miyagi, Chiba, Tokyo, Kanagawa, and Mie, Japan. Specimens collected by tympanotomy or myringotomy numbered 856, and 691 strains were isolated from 599 specimens. Of these, 219 isolates (31.7%) were identified as Streptococcus pneumoniae and 201 met study requirements. The most common serotype was 19F (52 isolates, 25.9%), followed by 6B (30 isolates, 14.9%) and 23F (24 isolates, 11.9%). Seven-valent vaccine serotype coverage was 62.7%. The percentage of PSSP was 40.3%, PISP 42.8%, and PRSP 16.9%, resistant strains (PISP and PRSP) combined accounted for 59.7%. Seven-valent vaccine serotype coverage for PISP was 80.2% and PRSP 82.4%. PBP gene mutation was observed in 175 isolates (87.1%), including 70 of gPISP (34.8%) and 105 of gPRSP (52.2%). Gene mutation induced by macrolides was found in 176 isolates (87.6%).
Subject(s)
Otitis Media, Suppurative/microbiology , Pneumonia, Pneumococcal/epidemiology , Streptococcus pneumoniae/classification , Acute Disease , Child, Preschool , Drug Resistance, Bacterial , Female , Humans , Infant , Macrolides/pharmacology , Male , Penicillin Resistance , Penicillin-Binding Proteins/genetics , Prospective Studies , Serotyping , Streptococcus pneumoniae/drug effects , Streptococcus pneumoniae/isolation & purificationABSTRACT
CONCLUSIONS: The proportion of drug-resistant bacteria was lower than previous reports. In children with acute otitis media (AOM), lower age, presence of multiple bacteria, and otitis media with effusion (OME) represented significant factors for recurrent AOM and the presence of drug-resistant bacteria. OBJECTIVE: Recently, the proportion of drug-resistant bacteria has been increasing in children with AOM. We studied the proportion of drug-resistant bacteria and background factors for detection of drug-resistant bacteria and recurrent AOM in a private clinic. SUBJECTS AND METHODS: Subjects comprised 170 patients <12 years old with AOM. Middle ear fluid was collected and pathogenic bacteria were identified. The following factors were considered: age, sex, use of antibiotics in the past 1 month, past history of recurrent AOM, presence of OME, and multiple bacteria of the three main strains (Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis). RESULTS: A total of 169 strains were detected in 77% of children with AOM. Drug-resistant bacteria comprised 44 of the 169 strains (26%). Lower age (p=0.001) and presence of multiple bacteria (p<0.001) represented significant factors for the presence of drug-resistant bacteria. OME was a significant factor for recurrent AOM (p<0.001).
