[Detection of Aspergillus spp. by real-time PCR in a murine model of pulmonary infection].

Abstract

OBJECTIVES:

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).

CONCLUSION:

Real-time PCR is a reliable procedure for the quantification and evaluation pulmonary infection due to A. fumigatus in animal models.

• Authors: María José Buitrago, Alicia Gómez-López, Emilia Mellado, Juan Luis Rodríguez-Tudela, Manuel Cuenca-Estrella
• Journal: Enfermedades infecciosas y microbiologia clinica (Enferm Infecc Microbiol Clin) Vol. 23 Issue 8 Pg. 464-8 (Oct 2005) ISSN: 0213-005X [Print] Spain
• Vernacular Title: Detección de Aspergillus spp. mediante PCR en tiempo real en un modelo murino de infección pulmonar.
• PMID: 16185559 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• DNA, Fungal

Topics

• Animals
• Aspergillosis, Allergic Bronchopulmonary (diagnosis)
• Aspergillus fumigatus (genetics)
• DNA, Fungal (analysis)
• Disease Models, Animal
• Male
• Mice
• Polymerase Chain Reaction