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首页> 外文期刊>Journal of Clinical Microbiology >Simple PCR-Based DNA Microarray System To Identify Human Pathogenic Fungi in Skin
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Simple PCR-Based DNA Microarray System To Identify Human Pathogenic Fungi in Skin

机译：简单的基于PCR的DNA微阵列系统，以识别皮肤中的人类致病真菌

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Fungal diseases in immunocompromised hosts pose significant threats to their prognoses. An accurate diagnosis and identification of the fungal pathogens causing the infection are critical to determine the proper therapeutic interventions, but these are often not achieved, due to difficulties with isolation and morphological identification. In an effort to ultimately carry out the simultaneous detection of all human pathogenic microbes, we developed a simple system to identify 26 clinically important fungi by using a combination of PCR amplification and DNA microarray assay (designated PCR-DM), in which PCR-amplified DNA from the internal transcribed spacer region of the rRNA gene was hybridized to a DNA microarray fabricated with species-specific probes sets using the Bubble Jet technology. PCR-DM reliably identified all 26 reference strains; hence, we applied it to cases of onychomycosis, taking advantage of the accessibility of tissue from skin. PCR-DM detected fungal DNA and identified pathogens in 92% of 106 microscopy-confirmed onychomycosis specimens. In contrast, culture was successful for only 36 specimens (34%), 3 of which had results inconsistent with the results of PCR-DM, but sequence analysis of the isolates proved that the PCR-DM result was correct. Thus, PCR-DM provides a powerful method to identify pathogenic fungi with high sensitivity and speed directly from tissue specimens, and this concept could be applied to other fungal or nonfungal infectious human diseases in less accessible anatomical sites.
机译：免疫受损宿主中的真菌疾病对其预后构成了重大威胁。对引起感染的真菌病原体进行准确的诊断和鉴定对于确定适当的治疗措施至关重要，但是由于分离和形态学鉴定的困难，这些措施常常无法实现。为了最终实现所有人类病原微生物的同时检测，我们开发了一种简单的系统，通过结合使用PCR扩增和DNA芯片测定（指定为PCR-DM）来鉴定26种临床上重要的真菌。使用Bubble Jet技术，将来自rRNA基因内部转录间隔区的DNA与用物种特异性探针组制造的DNA微阵列杂交。 PCR-DM可靠地鉴定了所有26种参考菌株。因此，我们利用皮肤组织的可及性将其应用于甲癣病例。 PCR-DM检测真菌DNA，并在106例经显微镜确认的灰指甲样本中92％识别出病原体。相比之下，仅36个标本（34％）的培养成功，其中3个的结果与PCR-DM的结果不一致，但是分离物的序列分析证明PCR-DM的结果是正确的。因此，PCR-DM提供了一种强有力的方法，可以直接从组织标本中以高灵敏度和高速度来鉴定病原真菌，并且该概念可以应用于难以接近的解剖部位的其他真菌或非真菌感染性人类疾病。

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