Centro Nacional de Microbiología, Instituto de Salud Carlos III, Madrid, Spain.
Med Mycol. 2012 Apr;50(3):270-5. doi: 10.3109/13693786.2011.604047. Epub 2011 Sep 12.
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.
建立并验证了一种用于检测镰刀菌属成员 DNA 的双路实时聚合酶链反应(RT-PCR)检测方法,该方法使用了两种侵袭性感染的小鼠模型。双路 RT-PCR 技术采用了针对真菌 rDNA 基因高度保守区的两个特异性分子信标探针。该技术的检测限为 10 fg DNA/μl 样本,特异性为 100%。在总共 48 份来自镰刀菌属感染的小鼠模型的样本中,肺组织的灵敏度为 93.9%,血清样本的灵敏度为 86.7%。相比之下,在总共 45 份 F. oxysporum 感染的小鼠模型样本中,肺组织的灵敏度为 87%,血清样本的灵敏度为 42.8%。该分子技术可能是一种可靠的方法,可用于动物模型中疾病的定量和评估,以及 Fusariosis 的临床诊断。