Lee Tracy, Cabrera Adriana, Kolehmainen Kathleen, Hird Trevor, Jorgensen Danielle, O'Dwyer Alan, Fornika Dan, KhunKhun Rupinder Kaur, Rodrigues Mabel, Prystajecky Natalie, Tyson John, Sekirov Inna, Zlosnik James E A
British Columbia Centre for Disease Control, Public Health Laboratory, Vancouver, BC V6Z R4R, Canada.
Department of Pathology and Laboratory Medicine, Faculty of Medicine, University of British Columbia, Vancouver, BC V6T 1Z4, Canada.
Trop Med Infect Dis. 2025 Jul 9;10(7):192. doi: 10.3390/tropicalmed10070192.
Evolving technologies available to clinical laboratories and laboratory-related updates to clinical guidelines both drive the need for clinical laboratories to keep their test menu updated and in line with current technological and clinical developments. Our laboratory has developed a targeted Illumina-based amplicon next-generation sequencing (NGS) assay to interrogate the 65 and 41) genes of spp. for the purposes of providing species-level ± subspecies-level identification of spp. organisms in clinical samples and genotypic predictions for inducible macrolide resistance (in the case of complex members). The developed assay demonstrated 100% sensitivity and specificity for and complex cultured organisms, 98% ID overall concordance relative to the available reference identification, and a nearly 60% "rescue" rate for primary samples that could not be identified using our previous method. There was 94.6% concordance between genotypic and phenotypic results for inducible macrolide resistance. The developed assay was successfully implemented in our clinical laboratory and has been accredited for clinical use.
临床实验室可获得的不断发展的技术以及临床指南中与实验室相关的更新，都促使临床实验室需要不断更新其检测项目，使其与当前的技术和临床发展保持一致。我们的实验室开发了一种基于Illumina的靶向扩增子新一代测序（NGS）检测方法，用于检测 spp. 的65个和41个基因，目的是对临床样本中的 spp. 生物体进行物种水平±亚种水平的鉴定，并对诱导型大环内酯耐药性进行基因型预测（对于 复合体成员而言）。所开发的检测方法对 和 复合体培养生物体显示出100%的敏感性和特异性，相对于可用的参考鉴定，总体鉴定一致性为98%，对于使用我们以前的方法无法鉴定的原始样本，“挽救”率接近60%。诱导型大环内酯耐药性的基因型和表型结果之间的一致性为94.6%。所开发的检测方法已在我们的临床实验室成功实施，并已获得临床使用认证。
