Identification of Mycobacterium Species by DNA Microarray Chip Method

• Published in: Journal of visualized experiments no. 220
• Main Authors: Zixuan, Chen, Jianming, Zhang, Yancheng, Jiang, Zhenglong, Lin, Xiaoting, Chen
• Format: Journal Article
• Language: English
• Published: United States 24.06.2025
• Subjects: DNA, Bacterial - analysis; DNA, Bacterial - genetics; Humans; Mycobacterium - classification; Mycobacterium - genetics; Mycobacterium - isolation & purification; Mycobacterium Infections - diagnosis; Mycobacterium Infections - microbiology; Oligonucleotide Array Sequence Analysis - methods; Polymerase Chain Reaction - methods
• DOI: 10.3791/68234

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.