Department of Pathology and Molecular Medicine, Hirosaki University Graduate School of Medicine, Zaifu-cho, Hirosaki, Japan.
Jpn J Infect Dis. 2010 May;63(3):188-91.
Mycobacterial infection of musculoskeletal tissue is a rare disease that may cause destruction of the tissues. Both Mycobacterium tuberculosis and non-tuberculous mycobacteria affect the tissues. Although surgical debridement and antibiotic therapy are required for the treatment, it is necessary to identify the causative species before selecting the antibiotics. However, it is difficult to identify the species in clinical samples from musculoskeletal tissue. In the current study, using pathological specimens, the causative species was identified by PCR amplification and direct sequencing of mycobacterial 16S rDNA containing a hypervariable region. Twelve cases of chronic granulomatous inflammation of musculoskeletal tissues were used for the study. DNA was extracted from paraffin sections, and mycobacterial 16S rDNA was amplified by PCR. The amplicons were obtained in 5 of 12 cases (41%), even in specimens in which the microorganism was only scarcely detected by using special stains. Direct sequencing of the amplified products presented high homology with M. tuberculosis in four cases and M. avium in one. Therefore, PCR-direct sequencing of 16S rDNA containing hypervariable region using pathological specimens is useful for the diagnosis and identification of causative species in mycobacterial infection of musculoskeletal tissues.
肌肉骨骼组织中的分枝杆菌感染是一种罕见的疾病，可能导致组织破坏。结核分枝杆菌和非结核分枝杆菌均可影响组织。尽管外科清创术和抗生素治疗是必需的，但在选择抗生素之前，有必要确定致病物种。然而，从肌肉骨骼组织的临床样本中鉴定物种是很困难的。在本研究中，使用病理标本，通过 PCR 扩增和直接测序含有高变区的分枝杆菌 16S rDNA 来鉴定致病物种。研究使用了 12 例肌肉骨骼组织慢性肉芽肿性炎症的病例。从石蜡切片中提取 DNA，通过 PCR 扩增分枝杆菌 16S rDNA。在 12 例病例中的 5 例（41%）中获得了扩增子，即使使用特殊染色法仅很少检测到微生物的标本也是如此。扩增产物的直接测序在 4 例中与结核分枝杆菌高度同源，在 1 例中与鸟分枝杆菌高度同源。因此，使用病理标本对含有高变区的 16S rDNA 进行 PCR-直接测序有助于诊断和鉴定肌肉骨骼组织分枝杆菌感染的致病物种。
