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应用基于双重实时定量 PCR 的方法检测在镰刀菌病的鼠模型中由茄病镰刀菌和非茄病镰刀菌种引起的侵袭性感染。

Detection of invasive infection caused by Fusarium solani and non-Fusarium solani species using a duplex quantitative PCR-based assay in a murine model of fusariosis.

机构信息

Centro Nacional de Microbiología, Instituto de Salud Carlos III, Madrid, Spain.

出版信息

Med Mycol. 2012 Apr;50(3):270-5. doi: 10.3109/13693786.2011.604047. Epub 2011 Sep 12.

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.

摘要

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。

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