Murine model of disseminated fusariosis: evaluation of the fungal burden by traditional CFU and quantitative PCR.
Systemic disease is the most severe clinical form of fusariosis, and the treatment involves a challenge due to the refractory response to antifungals. Treatment for murine Fusarium solani infection has been described in models that employ CFU quantitation in organs as a parameter of therapeutic efficacy. However, CFU counts do not precisely reproduce the amount of cells for filamentous fungi such as F. solani. In this study, we developed a murine model of disseminated fusariosis and compared the fungal burden with two methods: CFU and quantitative PCR. ICR and BALB/c mice received an intravenous injection of 1 × 10(7) conidia of F. solani per mouse. On days 2, 5, 7, and 9, mice from each mice strain were killed. The spleen and kidneys of each animal were removed and evaluated by qPCR and CFU determinations. Results from CFU assay indicated that the spleen and kidneys had almost the same fungal burden in both BALB/c and ICR mice during the days of the evaluation. In the qPCR assay, the spleen and kidney of each mouse strain had increased fungal burden in each determination throughout the entire experiment. The fungal load determined by the qPCR assay was significantly greater than that determined from CFU measurements of tissue. qPCR could be considered as a tool for quantitative evaluation of fungal burden in experimental disseminated F. solani infection.
Departamento de Microbiología, Facultad de Medicina, Universidad Autónoma de Nuevo León, Madero y Dr. Eduardo A. Pequeño s/n, Colonia Mitras Centro, 64460, Monterrey, NL, Mexico.
Gloria M
GM
González
Jazmín
J
Márquez
Rogelio de J
Rde J
Treviño-Rangel
José P
JP
Palma-Nicolás
Elvira
E
Garza-González
Luis A
LA
Ceceñas
J
J
Gerardo González
D000818:Animals; D015169:Colony Count, Microbial / Q000379:methods*; D004195:Disease Models, Animal; D060585:Fusariosis / Q000382:microbiology*; D005670:Fusarium / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D007668:Kidney / Q000382:microbiology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008813:Mice, Inbred ICR; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D013154:Spleen / Q000382:microbiology; D013997:Time Factors