Abstract
A duplex Real Time PCR (RT-PCR) assay for detecting DNA of members of the genus Fusarium has been developed and validated by using two mouse models of invasive infection. The duplex RT-PCR technique employed two specific molecular beacon probes targeting a highly conserved region of the fungal rDNA gene. This technique showed a detection limit of 10 fg DNA per μl of sample and a specificity of 100%. The sensitivity in a total of 48 samples from a murine model of Fusarium solani infection was 93.9% for lung tissues and 86.7% for serum samples. In comparison, the sensitivity in a total of 45 samples of a F. oxysporum murine model infection was 87% for lung tissues and 42.8% for serum samples. This molecular technique could be a reliable method for the quantification and the evaluation of the disease in animal models and for the clinical diagnosis of fusariosis.
Acknowledgements
This work was supported by Research Projects from Fundación Areces (MPY1397/04), from Instituto de Salud Carlos III (MPY1279/05), from Spanish Ministry of Science and Education (SAF2006-11627) and from Red Española de Investigación en Patología Infecciosa (REIPI). L. Bernal-Martinez has a research contract from REIPI (Red Española de Investigación en Patología Infecciosa, Project MPY 1022/07_1) and M V. Castelli had a research contract from AECI (Agencia Española de Cooperación Internacional).
Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.
This paper was first published online on Early Online on 7 September 2011.