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首页> 外文期刊>Journal of Clinical Microbiology >Development and Application of Real-Time PCR Assay for Quantification of Mycobacterium ulcerans DNA
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Development and Application of Real-Time PCR Assay for Quantification of Mycobacterium ulcerans DNA

机译：实时PCR检测定量分支杆菌DNA的开发与应用

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Buruli ulcer, an infection caused by Mycobacterium ulcerans, is, after tuberculosis and leprosy, the third most common mycobacterial disease. The mode of transmission of M. ulcerans is not exactly known, but since Buruli ulcer often occurs in focalized swampy areas, it is assumed that there is a reservoir of the pathogen in stagnant water. Buruli ulcer usually starts as a painless nodule and can lead to massive destruction of skin, subcutaneous tissue, and eventually muscle and bone. Currently the only recommended treatment is wide surgical excision. In this report we describe the development of a real-time PCR method for the quantification of M. ulcerans DNA (IS2404 TaqMan). The highly specific assay is based on the detection of the M. ulcerans specific insertion sequence IS2404. The IS2404 TaqMan assay turned out to be about 10 times more sensitive than the available conventional PCR-based diagnostic test. It is demonstrated that the IS2404 TaqMan assay is suitable for the quantitative assessment of the dissemination of the mycobacteria in Buruli ulcer lesions. Prototype results obtained with excised tissue from a patient with a late preulcerative Buruli ulcer lesion reconfirmed earlier histopathological findings indicating that tissue damage occurs far beyond the regions in which large numbers of mycobacteria are detectable. The IS2404 TaqMan assay should be a useful tool for both diagnosis and research into the pathology and mode of transmission of this still inadequately investigated mycobacterial disease.
机译：布鲁氏溃疡是由溃疡分枝杆菌引起的，是继结核和麻风病之后第三大最常见的分支杆菌病。 M的传输方式。溃疡还不是很清楚，但是由于布鲁氏溃疡经常发生在集中的沼泽地区，因此人们认为在死水中存在病原体的储藏库。布鲁氏溃疡通常从无痛结节开始，可导致皮肤，皮下组织的大量破坏，最终导致肌肉和骨骼的破坏。目前，唯一推荐的治疗方法是广泛的手术切除。在本报告中，我们描述了实时定量PCR定量M的方法的发展。溃疡DNA（IS2404 TaqMan）。高度特异性的测定是基于对 M的检测。溃疡的特异性插入序列是IS 2404 。结果证明，IS2404 TaqMan分析的灵敏度比可用的常规基于PCR的诊断测试高约10倍。结果表明，IS2404 TaqMan测定法适用于定量评估Buruli溃疡病变中分枝杆菌的分布。用溃疡性溃疡前溃疡晚期患者的切除的组织获得的原型结果再次证实了较早的组织病理学发现，表明组织损伤的发生范围远远超出了可检测到大量分枝杆菌的区域。 IS2404 TaqMan测定法应该是诊断和研究尚未充分研究的分枝杆菌疾病的病理学和传播方式的有用工具。

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