Identification of <em>Mycobacterium</em> Species by DNA Microarray Chip Method

DNA微阵列 微阵列 鉴定(生物学) 生物 DNA 分子生物学 微生物学 计算生物学 遗传学 基因 基因表达 植物
作者
Zixuan Chen,Jianming Zhang,Yancheng Jiang,Lin Zhenglong,Chen Xiaoting
出处
期刊:Journal of Visualized Experiments [MyJOVE]
卷期号: (220)
标识
DOI:10.3791/68234
摘要

This study presents a DNA microarray chip method designed for the accurate identification of Mycobacterium species. By leveraging asymmetric PCR amplification and hybridization principles, this method targets 17 common mycobacteria, including those in the Mycobacterium tuberculosis complex and various non-tuberculous mycobacteria. It is applicable to a wide range of clinical specimens like sputum, pus, bronchoalveolar lavage fluid, cerebrospinal fluid, and puncture fluid from patients suspected of mycobacterial diseases. The assay involves amplifying target sequences in the Mycobacterium genome through asymmetric PCR, followed by hybridization with the probes on the microarray chip. The unique probe arrangement (repeated 5x in a 12 row x 10 column microarray) and the use of control probes enhance its reliability. Clinical trials with 1,724 samples demonstrated high performance. The method achieved 100% clinical specificity, sensitivity, and overall concordance compared to sequencing results, except for two rare non-tuberculous mycobacterial samples beyond its detection scope. This DNA microarray chip method offers a significant improvement over traditional diagnostic techniques, shortening the diagnosis time and providing a more comprehensive detection, thus having great potential in the diagnosis and management of mycobacterial diseases.

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