通过16S核糖体DNA测序评估用于鉴定分枝杆菌的MicroSeq系统及其在常规临床分枝杆菌学实验室中的整合。
Evaluation of the MicroSeq system for identification of mycobacteria by 16S ribosomal DNA sequencing and its integration into a routine clinical mycobacteriology laboratory.
作者信息
Hall Leslie, Doerr Kelly A, Wohlfiel Sherri L, Roberts Glenn D
机构信息
Department of Laboratory Medicine and Pathology, Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota 55905, USA.
出版信息
J Clin Microbiol. 2003 Apr;41(4):1447-53. doi: 10.1128/JCM.41.4.1447-1453.2003.
An evaluation of the MicroSeq 500 microbial identification system by nucleic acid sequencing and the Mayo Clinic experience with its integration into a routine clinical laboratory setting are described. Evaluation of the MicroSeq 500 microbial identification system was accomplished with 59 American Type Culture Collection (ATCC) strains and 328 clinical isolates of mycobacteria identified by conventional and 16S ribosomal DNA sequencing by using the MicroSeq 500 microbial identification system. Nucleic acid sequencing identified 58 of 59 (98.3%) ATCC strains to the species level or to the correct group or complex level. The identification results for 219 of 243 clinical isolates (90.1%) with a distance score of <1% were concordant with the identifications made by phenotypic methods. The remaining 85 isolates had distance scores of >1%; 35 (41.1%) were identified to the appropriate species level or group or complex level; 13 (15.3%) were identified to the species level. All 85 isolates were determined to be mycobacterial species, either novel species or species that exhibited significant genotypic divergence from an organism in the database with the closest match. Integration of nucleic acid sequencing into the routine mycobacteriology laboratory and use of the MicroSeq 500 microbial identification system and Mayo Clinic databases containing additional genotypes of common species and added species significantly reduced the number of organisms that could not be identified by phenotypic methods. The turnaround time was shortened to 24 h, and results were reported much earlier. A limited number of species could not be differentiated from one another by 16S ribosomal DNA sequencing; however, the method provides for the identification of unusual species and more accurate identifications and offers the promise of being the most accurate method available.
本文描述了通过核酸测序对MicroSeq 500微生物鉴定系统的评估，以及梅奥诊所将其整合到常规临床实验室环境中的经验。使用MicroSeq 500微生物鉴定系统，对59株美国典型培养物保藏中心（ATCC）菌株和328株经传统方法及16S核糖体DNA测序鉴定的分枝杆菌临床分离株进行了MicroSeq 500微生物鉴定系统的评估。核酸测序将59株ATCC菌株中的58株（98.3%）鉴定到种水平或正确的组或复合体水平。243株临床分离株中，距离分数<1%的219株（90.1%）的鉴定结果与表型方法的鉴定结果一致。其余85株分离株的距离分数>1%；其中35株（41.1%）被鉴定到适当的种水平或组或复合体水平；13株（15.3%）被鉴定到种水平。所有85株分离株均被确定为分枝杆菌属物种，要么是新物种，要么是与数据库中最匹配的生物体表现出显著基因型差异的物种。将核酸测序整合到常规分枝杆菌学实验室，并使用MicroSeq 500微生物鉴定系统以及包含常见物种和新增物种的其他基因型的梅奥诊所数据库，显著减少了无法通过表型方法鉴定的生物体数量。周转时间缩短至24小时，结果报告得更早。通过16S核糖体DNA测序无法区分少数物种；然而，该方法可用于鉴定不寻常的物种并提供更准确的鉴定结果，有望成为可用的最准确方法。
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