Rapid identification of Klebsiella pneumoniae, Corynebacterium kutscheri, and Streptococcus pneumoniae using triplex polymerase chain reaction in rodents.
Exp Anim
; 62(1): 35-40, 2013.
Article
en En
| MEDLINE
| ID: mdl-23357944
ABSTRACT
Klebsiella pneumoniae, Corynebacterium kutscheri, and Streptococcus pneumoniae are important pathogens that cause respiratory infections in laboratory rodents. In this study, we used species-specific triplex PCR analysis to directly detect three common bacterial pathogens associated with respiratory diseases. Specific targets were amplified with conventional PCR using the tyrB gene from K. pneumoniae, gyrB gene from C. kutscheri, and ply gene from S. pneumoniae. Our primers were tested against purified DNA from another eleven murine bacteria to determine primer specificity. Under optimal PCR conditions, the triplex assay simultaneously yielded a 931 bp product from K. pneumoniae, a 540 bp product from C. kutscheri, and a 354 bp product from S. pneumoniae. The triplex assay detection thresholds for pure cultures were 10 pg for K. pneumoniae and S. pneumoniae, and 100 pg for C. kutscheri. All three bacteria were successfully identified in the trachea and lung of experimentally infected mice at the same time. Our triplex PCR method can be used as a useful method for detecting pathogenic bacterial infections in laboratory rodents.
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Base de datos:
MEDLINE
Asunto principal:
Infecciones del Sistema Respiratorio
/
Enfermedades de los Roedores
/
Streptococcus pneumoniae
/
Reacción en Cadena de la Polimerasa
/
Corynebacterium
/
Klebsiella pneumoniae
/
Animales de Laboratorio
/
Ratones Endogámicos BALB C
Tipo de estudio:
Diagnostic_studies
Límite:
Animals
Idioma:
En
Revista:
Exp Anim
Asunto de la revista:
MEDICINA VETERINARIA
Año:
2013
Tipo del documento:
Article