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播散性镰刀菌病的鼠模型:传统 CFU 和定量 PCR 评估真菌负担。

Murine model of disseminated fusariosis: evaluation of the fungal burden by traditional CFU and quantitative PCR.

机构信息

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.

出版信息

Mycopathologia. 2013 Oct;176(3-4):219-24. doi: 10.1007/s11046-013-9687-3. Epub 2013 Aug 13.

Abstract

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.

摘要

系统性疾病是最严重的镰刀菌病临床形式,由于对抗真菌药物的难治性反应,治疗具有挑战性。在采用器官 CFU 定量作为治疗效果参数的模型中,已经描述了对鼠类茄病镰刀菌感染的治疗。然而,CFU 计数并不能精确地再现丝状真菌(如茄病镰刀菌)的细胞数量。在这项研究中,我们开发了一种播散性镰刀菌病的鼠模型,并比较了两种方法的真菌负担:CFU 和定量 PCR。ICR 和 BALB/c 小鼠每只接受 1×10(7)个茄病镰刀菌分生孢子的静脉注射。在第 2、5、7 和 9 天,从每种小鼠中杀死一组动物。取出每个动物的脾脏和肾脏,并通过 qPCR 和 CFU 测定进行评估。CFU 测定结果表明,在评估期间,BALB/c 和 ICR 小鼠的脾脏和肾脏的真菌负担几乎相同。在 qPCR 测定中,在整个实验过程中,每个小鼠品系的脾脏和肾脏的真菌负担在每次测定中均增加。qPCR 测定的真菌负荷明显大于组织 CFU 测量的真菌负荷。qPCR 可以被认为是实验性播散性茄病镰刀菌感染中定量评估真菌负担的工具。

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