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首页> 美国卫生研究院文献>Journal of Clinical Microbiology >PCR Detection of DNA Specific for Trichosporon Species in Serum of Patients with Disseminated Trichosporonosis
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PCR Detection of DNA Specific for Trichosporon Species in Serum of Patients with Disseminated Trichosporonosis

机译：弥散性毛孢子菌病患者血清中毛孢子菌种特异DNA的PCR检测

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Deep-seated trichosporonosis is a lethal opportunistic infection that disseminates rapidly and widely in immunocompromised patients, and early diagnosis is crucial for the treatment of this infection. We developed a novel nested-PCR assay that detects DNA specific for clinically important strains of Trichosporon in serum samples from patients with disseminated trichosporonosis. In this assay, two sets of oligonucleotide primers were derived from the sequence of 26S rRNA genes of Trichosporon asahii. The specific fragment was amplified from T. asahii and T. mucoides, but not from other microorganisms, including some other basidiomycetous fungi (Cryptococcus, Malassezia, Rhodotorula, and Sporobolomyces). Target DNA was detected by the nested PCR with as little as 5 fg of the extracted DNA of T. asahii. In a study using 11 clinical samples, the specific fragment was detected by the nested PCR in 64% (7 of 11) of sera from patients with histologically diagnosed disseminated trichosporonosis, while glucuronoxylomannan antigen was detected in only 54% (6 of 11) of the samples. Our new nested-PCR assay using serum samples can be performed repeatedly throughout the course of the disease. In addition, not only can it be used for early diagnosis of trichosporonosis, but it may also be beneficial for monitoring its progress or response to therapy.
机译：深层毛滴虫病是一种致命的机会性感染，可在免疫功能低下的患者中广泛传播，而早期诊断对于治疗这种感染至关重要。我们开发了一种新型的巢式PCR检测技术，该检测方法可检测来自散发性毛孢子虫病患者血清样品中特异于临床上重要的毛孢子菌菌株的DNA。在该测定法中，从天毛粉孢菌的26S rRNA基因的序列衍生出两组寡核苷酸引物。特异片段是从拟南芥和粘液虫中扩增而来的，而不是从其他微生物中扩增而来的，包括一些其他担子菌真菌（隐球菌，马拉色菌，红球菌和孢子菌）。通过巢式PCR检测目标DNA，并提取少至5 fg的T. asahii DNA。在一项使用11个临床样品的研究中，通过巢式PCR在组织学诊断为弥散型毛滴虫病患者中64％（11个中的7个）血清中检测到了特定片段，而在其中的54％（11个中的6个）中检测到了葡萄糖醛酸甘露聚糖抗原。样本。我们新的使用血清样品的巢式PCR检测可以在整个疾病过程中重复进行。此外，它不仅可以用于早期诊断毛状孢子虫病，而且对监测其进展或对治疗的反应也可能是有益的。

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