Balasingham Seetha V, Davidsen Tonje, Szpinda Irena, Frye Stephan A, Tønjum Tone
Centre for Molecular Biology and Neuroscience, Institute of Microbiology, University of Oslo, Norway.
Mol Diagn Ther. 2009;13(3):137-51. doi: 10.1007/BF03256322.
The processing of clinical specimens in the mycobacterial diagnostic laboratory has undergone remarkable improvements during the last decade. While microscopy and culture are still the major backbone for laboratory diagnosis of tuberculosis on a worldwide basis, new methods including molecular diagnostic tests have evolved over the last two decades. The majority of molecular tests have been focused on (i) detection of nucleic acids, both DNA and RNA, that are specific to Mycobacterium tuberculosis, by amplification techniques such as polymerase chain reaction (PCR); and (ii) detection of mutations in the genes that are associated with resistance to antituberculosis drugs by sequencing or nucleic acid hybridization. Recent developments in direct and rapid detection of mycobacteria, with emphasis on M. tuberculosis species identification by 16S rRNA gene sequence analysis or oligohybridization and strain typing, as well as detection of drug susceptibility patterns, all contribute to these advances. Generally, the balance between genome instability and genome maintenance as the basis for evolutionary development, strain diversification and resistance development is important, because it cradles the resulting M. tuberculosis phenotype. At the same time, semi-automated culture systems have contributed greatly to the increased sensitivity and reduced turnaround time in the mycobacterial analysis of clinical specimens. Collectively, these advances are particularly important for establishing the diagnosis of tuberculosis in children. More basic and operational research to appraise the impact and cost effectiveness of new diagnostic technologies must, however, be carried out. Furthermore, the design and quality of clinical trials evaluating new diagnostics must be improved to allow clinical and laboratory services that would provide rapid response to test results. Thus, important work remains before the new diagnostic tools can be meaningfully integrated into national tuberculosis control programs of high-burden countries.
在过去十年中，分枝杆菌诊断实验室中临床标本的处理有了显著改进。虽然显微镜检查和培养仍是全球结核病实验室诊断的主要支柱，但在过去二十年中出现了包括分子诊断测试在内的新方法。大多数分子测试集中于：（i）通过聚合酶链反应（PCR）等扩增技术检测结核分枝杆菌特有的核酸，包括DNA和RNA；（ii）通过测序或核酸杂交检测与抗结核药物耐药性相关基因中的突变。分枝杆菌直接快速检测的最新进展，重点是通过16S rRNA基因序列分析或寡核苷酸杂交进行结核分枝杆菌菌种鉴定和菌株分型，以及检测药物敏感性模式，都促成了这些进步。一般来说，基因组不稳定性与基因组维持之间的平衡作为进化发展、菌株多样化和耐药性发展的基础很重要，因为它孕育了最终的结核分枝杆菌表型。同时，半自动培养系统极大地提高了临床标本分枝杆菌分析的灵敏度并缩短了周转时间。总体而言，这些进展对儿童结核病的诊断尤为重要。然而，必须开展更多基础和操作性研究以评估新诊断技术的影响和成本效益。此外，评估新诊断方法的临床试验的设计和质量必须改进，以便临床和实验室服务能够对检测结果做出快速反应。因此，在新诊断工具能够有意义地纳入高负担国家的国家结核病控制规划之前，仍有重要工作要做。
