National Reference Laboratory for Mycobacteria, National Institute of Tuberculosis, Pulmonary Diseases and Thoracic Surgery, 1 Vysne Hagy St., 059 84, Vysoke Tatry, Slovakia,
Adv Exp Med Biol. 2015;835:15-22. doi: 10.1007/5584_2014_28.
Tuberculosis currently belongs to rare respiratory diseases in Slovakia. However, the emergence and spread of multi-drug resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) are major challenges for global tuberculosis control, since the treatment of resistant forms creates both medical and financial problems. Cultivation methods of diagnosis are time-consuming, many times exceeding the time of the initial phase of tuberculosis treatment. Therefore, in the presented study we compared the standard procedures, based on the cultivation of mycobacteria and subsequent drug susceptibility testing to antituberculotics, with molecular-genetic methods using PCR diagnostic kits. The molecular-genetic testing enables to obtain direct and fast evidence of Mycobacterium tuberculosis, with genomic verification of resistance to the most important anti-tuberculosis drugs - isoniazid and rifampicin in MDR-TB, and ethambutol, aminoglycosides, and fluoroquinolones in XDR-TB. In 2012-2013, we confirmed 19 cases of drug-resistant tuberculosis in Slovakia. The resistance to rifampicin was confirmed in all strains with both methods. In two cases, the molecular-genetic testing did not show resistance to isoniazid, as confirmed by conventional cultivation. Furthermore, two strains demonstrating susceptibility in conventional microbiological testing to ethambutol and five strains to fluoroquinolones were verified as actually being resistant using a PCR method. Rapid diagnosis and identification of MDR-TB or XDR-TB strains using molecular-genetic testing is an essential tool for the timely and appropriate drug treatment and prevention of spread of drug resistant strains.
结核病目前在斯洛伐克属于罕见的呼吸道疾病。然而，耐多药结核病（MDR-TB）和广泛耐药结核病（XDR-TB）的出现和传播是全球结核病控制的主要挑战，因为耐药形式的治疗既带来了医疗问题，也带来了财政问题。诊断的培养方法耗时，多次超过结核病治疗的初始阶段。因此，在本研究中，我们将基于分枝杆菌培养和随后的抗结核药物敏感性测试的标准程序与使用 PCR 诊断试剂盒的分子遗传学方法进行了比较。分子遗传学检测能够直接快速地获得结核分枝杆菌的证据，并对 MDR-TB 中的异烟肼和利福平以及 XDR-TB 中的乙胺丁醇、氨基糖苷类和氟喹诺酮类等最重要的抗结核药物的耐药性进行基因组验证。2012-2013 年，我们在斯洛伐克确认了 19 例耐药结核病病例。两种方法均证实了所有菌株对利福平的耐药性。在两种情况下，分子遗传学检测未显示对异烟肼的耐药性，这与常规培养所证实的一致。此外，两种在常规微生物学检测中对乙胺丁醇表现出敏感性的菌株和五种对氟喹诺酮类药物表现出敏感性的菌株，通过 PCR 方法证实实际上具有耐药性。使用分子遗传学检测快速诊断和鉴定 MDR-TB 或 XDR-TB 菌株是及时进行适当药物治疗和防止耐药菌株传播的重要工具。
