Scedosporium apiospermum and Scedosporium prolificans are fungal pathogens that can cause severe human infections, including disseminated mycosis in immunocompromised patients. Two real-time PCR (RT-PCR) assays for the diagnosis of these species were developed and validated for the classification of clinical strains and for the detection of DNA in clinical samples by use of a murine model of invasive infection. A total of 14 clinical strains and 141 samples, including blood, serum, and lung samples from infected CD1 mice, were analyzed. Each RT-PCR methodology used a species-specific molecular beacon probe targeting a highly conserved region of the fungal ribosomal DNA gene. Results showed 100% specificity and a detection limit of 10 fg of DNA for both assays. The sensitivities for the S. prolificans-specific PCR assay were 100% for cultured clinical strains, 95.5% for lung tissues, 85% for serum, and 83.3% for blood. For S. apiospermum, the sensitivities were 100% for clinical strains and 97.2%, 81.8%, and 54.5% for lung tissues, serum, and blood, respectively. Both techniques can be useful for clinical diagnosis, and further studies are warranted.
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机译：臭皮孢孢子菌和产多孢菌孢子菌是真菌病原体，可引起严重的人类感染，包括免疫受损患者的弥散性真菌病。开发了两种用于诊断这些物种的实时PCR（RT-PCR）方法，并通过使用侵袭性小鼠模型对临床菌株的分类和临床样品中DNA的检测进行了验证。共分析了14种临床菌株和141个样品，包括来自感染CD1小鼠的血液，血清和肺样品。每种RT-PCR方法都使用针对真菌核糖体DNA基因高度保守区域的物种特异性分子信标探针。结果表明，两种测定的特异性均为100％，DNA的检出限为10 fg。 S. prolificans特异性PCR检测法对培养的临床菌株的敏感性为100％，对肺组织的敏感性为95.5％，对血清的敏感性为85％，对血液的敏感性为83.3％。对于api.perospermum，临床菌株的敏感性分别为100％，对肺组织，血清和血液的敏感性分别为97.2％，81.8％和54.5％。两种技术都可用于临床诊断，因此有必要进行进一步的研究。
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