title: Detection of Streptococcus pneumoniae and identification of pneumococcal serotypes by real-time polymerase chain reaction using blood samples from Italian children ≤ 5 years of age with community-acquired pneumonia.
abstract: Streptococcus pneumoniae is a leading cause of severe life-threatening infections. Laboratory identification and serotyping of this pathogens is desirable to monitor vaccine impact and coverage; however, especially in pediatric patients, the yield of traditional microbiological diagnostic procedures can be very low. The aim of this study was to develop real-time polymerase chain reaction (PCR)-based assays to be performed directly on blood samples to identify the most common capsular serotypes causing pneumonia in Italian children (≤ 5 years of ages) after the introduction of the 7-valent conjugate vaccine. Our real-time PCR-based assays showed high sensitivity (at least 35 fg of pneumococcal DNA), and they were validated with 49 well-characterized pneumococcal isolates, 8 nonpneumococcal isolates, 13 simulated blood clinical samples loaded with S. pneumoniae of known serotypes, and 46 blood clinical samples. All the strains tested and the simulated blood clinical samples were correctly typed by the technique. Real-time PCR allowed serotyping in 37/46 children ≤ 5 years of age (80.4%) in whom pneumonia was diagnosed in four Italian hospitals. Non-PCV7 serotypes accounted for at least 47.8% (22/46) of cases, serotype 19A being the most common (34.7%, 16/46). Although, it is not known at present whether the incidence of 19A serotype is attributable to the use of PCV7 only, expanding pneumococcal serotype coverage has clearly the potential to prevent a larger number of pneumonias in Italian children less than ≤ 5 years of age. Molecular methods are of increasing importance in the diagnosis of pneumococcal pneumonia and in monitoring serotype distribution and replacement.
authors.affiliation: Dipartimento Interdisciplinare di Scienze Chiaurgiche e Diagnostiche Integrate, Sezione di Microbiologia, University of Genoa, Genoa, Italy. anna.marchese@unige.it
authors.forename: Anna
authors.initials: A
authors.lastname: Marchese
authors.forename: Susanna
authors.initials: S
authors.lastname: Esposito
authors.forename: Erika
authors.initials: E
authors.lastname: Coppo
authors.forename: Giovanni A
authors.initials: GA
authors.lastname: Rossi
authors.forename: Alberto
authors.initials: A
authors.lastname: Tozzi
authors.forename: Mariateresa
authors.initials: M
authors.lastname: Romano
authors.forename: Liviana
authors.initials: L
authors.lastname: Da Dalt
authors.forename: Gian Carlo
authors.initials: GC
authors.lastname: Schito
authors.forename: Nicola
authors.initials: N
authors.lastname: Principi
mesh_terms: D000328:Adult; D002675:Child, Preschool; D017714:Community-Acquired Infections / Q000097:blood / Q000382:microbiology* / Q000517:prevention & control; D000069443:Heptavalent Pneumococcal Conjugate Vaccine; D006801:Humans; D015994:Incidence; D007558:Italy; D011008:Pneumococcal Infections / Q000097:blood / Q000382:microbiology* / Q000517:prevention & control; D022242:Pneumococcal Vaccines / Q000008:administration & dosage; D011018:Pneumonia, Pneumococcal / Q000097:blood / Q000382:microbiology* / Q000517:prevention & control; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D012703:Serotyping / Q000379:methods; D013296:Streptococcus pneumoniae / Q000145:classification / Q000235:genetics*; D018074:Vaccines, Conjugate / Q000008:administration & dosage
chemical_list: D000069443:Heptavalent Pneumococcal Conjugate Vaccine; D022242:Pneumococcal Vaccines; D018074:Vaccines, Conjugate