Troesch A, Nguyen H, Miyada C G, Desvarenne S, Gingeras T R, Kaplan P M, Cros P, Mabilat C
bioMerieux, 69280 Marcy-L'Etoile, France.
J Clin Microbiol. 1999 Jan;37(1):49-55. doi: 10.1128/JCM.37.1.49-55.1999.
Species identification within the genus Mycobacterium and subsequent antibiotic susceptibility testing still rely on time-consuming, culture-based methods. Despite the recent development of DNA probes, which greatly reduce assay time, there is a need for a single platform assay capable of answering the multitude of diagnostic questions associated with this genus. We describe the use of a DNA probe array based on two sequence databases: one for the species identification of mycobacteria (82 unique 16S rRNA sequences corresponding to 54 phenotypical species) and the other for detecting Mycobacterium tuberculosis rifampin resistance (rpoB alleles). Species identification or rifampin resistance was determined by hybridizing fluorescently labeled, amplified genetic material generated from bacterial colonies to the array. Seventy mycobacterial isolates from 27 different species and 15 rifampin-resistant M. tuberculosis strains were tested. A total of 26 of 27 species were correctly identified as well as all of the rpoB mutants. This parallel testing format opens new perspectives in terms of patient management for bacterial diseases by allowing a number of genetic tests to be simultaneously run.
分枝杆菌属内的菌种鉴定以及后续的药敏试验仍然依赖耗时的基于培养的方法。尽管最近开发出了DNA探针，大大缩短了检测时间，但仍需要一个单一平台检测方法来回答与该属相关的众多诊断问题。我们描述了一种基于两个序列数据库的DNA探针阵列的应用：一个用于分枝杆菌的菌种鉴定（对应54个表型菌种的82个独特的16S rRNA序列），另一个用于检测结核分枝杆菌对利福平的耐药性（rpoB等位基因）。通过将从细菌菌落产生的荧光标记的扩增遗传物质与阵列杂交来确定菌种鉴定或利福平耐药性。对来自27个不同菌种的70株分枝杆菌分离株和15株耐利福平的结核分枝杆菌菌株进行了检测。27个菌种中的26个以及所有rpoB突变体均被正确鉴定。这种平行检测形式通过允许同时进行多项基因检测，为细菌性疾病的患者管理开辟了新的前景。
