Rapid pathogen identification using a novel microarray-based assay with purulent meningitis in cerebrospinal fluid.
Rapid pathogen identification using a novel microarray-based assay with purulent meningitis in cerebrospinal fluid.
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使用基于微阵列的新型检测方法快速鉴定脑脊液中化脓性脑膜炎的病原体
DOI：
10.1038/s41598-018-34051-0
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发表时间：
2018-10-29
影响因子：
4.6
通讯作者：
Wang Z
中科院分区：
文献类型：
--
作者：
Hou Y;Zhang X;Hou X;Wu R;Wang Y;He X;Wang L;Wang Z
为了提高化脓性脑膜炎脑脊液中病原菌的诊断水平，我们建立了同时检测和鉴定7种靶菌的DNA芯片技术。从24例化脓性脑膜炎(或疑似化脓性脑膜炎)的脑脊液标本中提取DNA。选择每个病原体的特异性基因作为扩增目标，进行聚合酶链式反应(PCR)，用荧光染料标记，与芯片上的寡核苷酸探针杂交。在非靶标细菌中未出现明显的交叉杂交荧光信号。DNA芯片检测阳性率为87.5%(21/24)，脑脊液培养阳性率为58.3%(14/24)。其中58.3%(14/24)的患者经细菌培养证实为化脓性脑膜炎，37.5%(9/24)的患者经细菌培养未证实但经临床诊断和DNA芯片证实。基因芯片检测到5例多种细菌感染。此外，对基因拷贝数进行了检测，其灵敏度为3.5 × 10 1拷贝/μL。结果表明，针对脑脊液标本中病原体的基因芯片技术比传统的培养方法具有更好的特异性、准确性和敏感性。基因芯片技术是快速检测更多的靶标病原体和确定菌株亚型的有效工具，可以消除不同个体化脓性脑膜炎的影响，及时诊断和治疗。
In order to improve the diagnosis of pathogenic bacteria in cerebrospinal fluid (CSF) with purulent meningitis, we developed a DNA microarray technique for simultaneous detection and identification of seven target bacterium. DNA were extracted from 24 CSF samples with purulent meningitis (or suspected purulent meningitis). The specific genes of each pathogen were chosen as the amplification target, performed the polymerase chain reaction (PCR), labeled with a fluorescence dye, and hybridized to the oligonucleotide probes on the microarray. There is no significant cross-hybridization fluorescent signal occurred in untargeted bacteria. There were 87.5% (21/24) positive results in DNA microarray compared with the 58.3% (14/24) of the CSF culture test. Of which 58.3% (14/24) of the patients with culture-confirmed purulent meningitis, 37.5% (9/24) patients who were not confirmed by culture test but were demonstrated by the clinical diagnosis and DNA microarray. Multiple bacterial infections were detected in 5 cases by the microarray. In addition, the number of gene copies was carried out to determine the sensitivity of this technique, which was shown to be 3.5 × 101copies/μL. The results revealed that the microarray technique which target pathogens of the CSF specimen is better specificity, accuracy, and sensitivity than traditional culture method. The microarray method is an effective tool for rapidly detecting more target pathogens and identifying the subtypes of strains which can eliminate the impact of the different individuals with purulent meningitis for prompt diagnosis and treatment.
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影响因子：
4.4
作者：
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通讯作者：
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作者：
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