[Detection of Aspergillus spp. by real-time PCR in a murine model of pulmonary infection].
OBJECTIVE
Assessment of a real-time PCR technique for the detection and quantification of fungal DNA in a murine model of pulmonary aspergillosis.


METHODS
Male ICR specific pathogen-free mice were used in the studies. The animals were divided into groups: immunosuppressed and intranasally inoculated with various inoculum sizes (10(6), 10(5), 10(4), and 10(3) conidia/mL) of a clinical isolate of Aspergillus fumigatus. When symptoms of pulmonary aspergillosis were detected, the mice were killed and the lungs removed for culture and real-time PCR determination. The PCR reactions used primers that amplified a region of Aspergillus spp. ribosomal DNA. Survival time per experimental group was calculated and correlation coefficients with inoculum size, colony counts and PCR results were determined.


RESULTS
Average survival time was significantly associated with the size of the inoculum. Pulmonary colony count was positive for 90% of the infected mice, but there was no statistical relationship between count values and either survival time or inoculum size. Real-time PCR was positive in 100% of the animals and was significantly associated with survival time and inoculum size (p < 0.01).


CONCLUSIONS
Real-time PCR is a reliable procedure for the quantification and evaluation pulmonary infection due to A. fumigatus in animal models.
Buitrago
María José
MJ
Servicio de Micología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Madrid, Spain.
Gómez-López
Alicia
A
Mellado
Emilia
E
Rodríguez-Tudela
Juan Luis
JL
Cuenca-Estrella
Manuel
M
D000818:Animals; D001229:Aspergillosis, Allergic Bronchopulmonary / Q000175:diagnosis*; D001232:Aspergillus fumigatus / Q000235:genetics*; D004271:DNA, Fungal / Q000032:analysis*; D004195:Disease Models, Animal; D008297:Male; D051379:Mice; D016133:Polymerase Chain Reaction
D004271:DNA, Fungal