Development of a new real-time PCR system for simultaneous detection of bacteria and fungi in pathological samples.
Development of a new real-time PCR system for simultaneous detection of bacteria and fungi in pathological samples.
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开发新型实时PCR系统，用于同时检测病理样本中的细菌和真菌。
DOI：
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发表时间：
2015
影响因子：
1.4
通讯作者：
H. Katano
中科院分区：
文献类型：
--
作者：
Hitomi Fukumoto;Yuko Sato;H. Hasegawa;H. Saeki;H. Katano
利用实时聚合酶链反应（PCR）技术建立了一种新的同时检测病理标本中病原菌和真菌的方法。该系统被命名为“多微生物实时PCR”，具有在96孔板格式中同时检测68种细菌和9种真菌的潜力。所有探针引物组设计为使用福尔马林固定的石蜡包埋的样品作为输入产生小于210 bp的扩增子。针对从特定病原体的纯培养物中提取的DNA测试每个探针引物组的特异性和灵敏度。多微生物实时PCR系统揭示了10名获得性免疫缺陷综合征患者尸检时收集的肺部样本中的微生物感染概况。金黄色葡萄球菌是检测到的最常见的微生物（n=8），但具有低拷贝数。高拷贝数的铜绿假单胞菌检测肺脓肿样本（n=6）。屎肠球菌（n=6）、脑膜炎败血症嗜酸金杆菌（n=4）和白色念珠菌（n=4）也经常检出。此外，在1例肺炎病例中检测到结核分枝杆菌潜伏感染。总之，这种多微生物实时PCR系统可以用于检测细菌和真菌的病理标本从不确定的诊断患者。
A novel system for simultaneous detection of pathogenic bacteria and fungi in pathological samples was developed using a real-time polymerase chain reaction (PCR) system. This system, designated the "multi-microbial real-time PCR", has the potential to simultaneously detect 68 bacterial and 9 fungal species in a 96-well plate format. All probe-primer sets were designed to produce amplicons smaller than 210 bp using formalin-fixed paraffin-embedded samples as input. The specificity and sensitivity of each probe-primer set were tested against DNA extracted from pure cultures of specific pathogens. The multi-microbial real-time PCR system revealed profiles of microorganism infection in lung samples collected at autopsy from 10 patients with acquired immunodeficiency syndrome. Staphylococcus aureus was the most common microbe detected (n=8), but with low copy numbers. High copy numbers of Pseudomonas aeruginosa were detected in the lung samples with abscess (n=6). Enterococcus faecium (n=6), Elizabethkingia meningoseptica (n=4), and Candida albicans (n=4) were also frequently detected. In addition, a latent infection of Mycobacterium tuberculosis was detected in one case of pneumonia. In conclusion, this multi-microbial real-time PCR system can be useful for detecting bacteria and fungi in pathological specimens from patients with uncertain diagnoses.
DOI：
10.1073/pnas.95.25.14863
发表时间：
1998-12-08
影响因子：
11.1
作者：
Eisen, MB;Spellman, PT;Botstein, D
通讯作者：
Botstein, D