Detection of Blastomyces dermatitidis and Histoplasma capsulatum from Culture Isolates and Clinical Specimens by Use of Real-Time PCR
Detection of Blastomyces dermatitidis and Histoplasma capsulatum from Culture Isolates and Clinical Specimens by Use of Real-Time PCR
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DOI：
10.1128/jcm.00673-11
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发表时间：
2011-09-01
影响因子：
9.4
通讯作者：
Wengenack, Nancy L.
中科院分区：
文献类型：
--
作者：
Babady, N. Esther;Buckwalter, Seanne P.;Wengenack, Nancy L.
皮肤芽生菌和荚膜组织胞浆菌是二型真菌，经常引起自限性呼吸道感染。然而，它们也可能导致严重的传播性疾病，这取决于接触生物体的程度和宿主的免疫状况。此外，由这些真菌引起的感染患者可能具有非常相似的临床表现。虽然微生物培养是检测这些病原体的标准方法，但它们的恢复可能需要数天至数周，并且培养物的操作对实验室人员构成重大安全危害。因此，本研究的目的是设计一种快速，实时PCR检测和区分B。dermativitis和H. capsulatum从培养分离株和直接从临床标本。针对B的组氨酸激酶和甘油醛-3-磷酸脱氢酶基因设计引物和荧光共振能量转移杂交探针。dermativitis和H. capsulatum，分别。测定两种真菌的分析灵敏度均为100拷贝/μ l。从培养分离株中，该试验证明了B的100%特异性和100%灵敏度。特异性为100%，敏感性为94%。胶囊。直接检测797例临床标本，B的特异性和敏感性分别为99%和86%。皮肤感染率分别为100%和73%。capsulatum与培养结果进行比较。这种实时PCR检测方法为B的检测提供了一种快速的方法。dermativitis和H. capsulatum从培养分离株和直接从临床标本。
Blastomyces dermatitidis and Histoplasma capsulatum are dimorphic fungi that often cause self-limited respiratory infections. However, they may also cause severe disseminated disease, depending on the level of the exposure to the organism and the host immune status. In addition, patients with infections caused by these fungi may have very similar clinical presentations. Although microbiologic culture is a standard method for detecting these pathogens, their recovery may require days to weeks, and the manipulation of cultures presents a significant safety hazard to laboratory personnel. Therefore, the goal of this study was to design a rapid, real-time PCR assay to detect and differentiate B. dermatitidis and H. capsulatum from culture isolates and directly from clinical specimens. Primers and fluorescence resonance energy transfer hybridization probes were designed to target the histidine kinase and glyceraldehyde-3-phosphate dehydrogenase genes of B. dermatitidis and H. capsulatum, respectively. The analytical sensitivity of the assay was determined to be 100 copies/mu l for both fungi. From culture isolates, the assay demonstrated 100% specificity and 100% sensitivity for B. dermatitidis and 100% specificity and 94% sensitivity for H. capsulatum. Detection directly from 797 clinical specimens demonstrated specificities and sensitivities of 99% and 86% for B. dermatitidis and 100% and 73% for H. capsulatum compared with the results for culture. This real-time PCR assay provides a rapid method for the detection of B. dermatitidis and H. capsulatum from culture isolates and directly from clinical specimens.