Molecular Diagnosis of Streptococcus pneumoniae in Middle Ear Fluids from Children with Otitis Media with Effusion
Pediatric Infection & Vaccine
;
: 106-112, 2015.
Artículo
en Inglés
| WPRIM
| ID: wpr-171626
ABSTRACT
PURPOSE:
The long-term administration of antibiotics interferes with bacterial culture in the middle ear fluids (MEFs) of young children with otitis media with effusion (OME). The purpose of this study is to determine whether molecular diagnostics can be used for rapid and direct detection of the bacterial pathogen in culture-negative MEFs.METHODS:
The specificity and sensitivity of both polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) to the lytA gene of Streptococcus pneumoniae were comparatively tested and then applied for pneumococcal detection in the clinical MEFs.RESULTS:
The detection limit of the PCR assay was approximately 10(4) colony forming units (CFU), whereas that of LAMP was less than 10 CFU for the detection of S. pneumoniae. Both PCR and LAMP did not amplify nucleic acid at over 10(6) CFU of H. influenzae or M. catarrhalis, both of which were irrelevant bacterial species. Of 22 culture-negative MEFs from children with OME, LAMP positivity was found in twelve MEFs (54.5%, 12/22), only three of which were PCR-positive (25%, 3/12). Our results showed that the ability of LAMP to detect pneumococcal DNA is over four times higher than that of PCR (P<0.01).CONCLUSIONS:
As a high-resolution tool able to detect nucleic acid levels equivalent to <10 CFU of S. pneumoniae in MEFs without any cross-reaction with other pathogens, lytA-specific LAMP may be applied for diagnosing pneumococcus infection in OME as well as evaluating the impact of a pneumococcal conjugate vaccine against OME.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Otitis
/
Otitis Media
/
Neumonía
/
Células Madre
/
Streptococcus
/
Streptococcus pneumoniae
/
Otitis Media con Derrame
/
ADN
/
Reacción en Cadena de la Polimerasa
/
Sensibilidad y Especificidad
Tipo de estudio:
Estudio diagnóstico
Límite:
Niño
/
Humanos
Idioma:
Inglés
Revista:
Pediatric Infection & Vaccine
Año:
2015
Tipo del documento:
Artículo
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