用于临床微生物全基因组测序测定的商用自动文库制备系统的性能评估。

IF 3.4 3区 医学 Q1 PATHOLOGY Journal of Molecular Diagnostics Pub Date : 2024-08-01 DOI:10.1016/j.jmoldx.2024.05.006
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引用次数: 0

摘要

下一代测序(NGS)已被证明在疾病管理方面具有临床用途,同时也是基因组监测的重要工具。目前,围绕其实施的障碍,即复杂而苛刻的分析工作流程,阻碍了其在许多实验室的广泛应用。为了应对这一挑战,加州大学洛杉矶分校分子微生物学与病原体基因组学(MMPG)实验室评估了 Tecan MagicPrep NGS 系统与 Illumina Nextera DNA Flex 文库制备系统的性能,前者是用于临床全基因组测序(WGS)检测的商用自动化文库制备解决方案。我们利用 35 种独特的生物(28 种细菌和 7 种真菌)进行了各种临床应用,包括微生物鉴定和基因组特征描述,比较了所制备文库的数量和质量、所得序列以及总体结果的一致性。我们还评估了其实施对实验室工作流程的影响。MagicPrep NGS 产生的文库浓度更高,体积更小,摩尔浓度也相应更高。然而,序列的质量指标没有显示出对总体结果的显著影响,与参考方法的一致性达到 100%。重要的是,工作流程分析表明,每次运行可减少 5 小时的操作时间,灵活性更高。我们的评估研究表明,MagicPrep NGS 的性能与 Nextera DNA Flex 相当,而且还能提高工作流程效率,减少进行常规临床微生物 WGS 检测的人力。
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Performance Evaluation of a Commercial Automated Library Preparation System for Clinical Microbial Whole-Genome Sequencing Assays

Next-generation sequencing (NGS) has proven clinical utility on disease management and serves as an important tool for genomic surveillance. Currently, hurdles surrounding its implementation, namely the complex and demanding analytical workflows, have impeded its widespread use in many laboratories. To address this challenge, the UCLA Molecular Microbiology and Pathogen Genomics Laboratory evaluated the performance of the Tecan MagicPrep NGS system, a commercial automated solution for library preparation for clinical whole-genome sequencing assays, against the Illumina Nextera DNA Flex Library Prep. Using 35 unique organisms (28 bacteria and 7 fungi) for various clinical applications, including microbial identification and genomic characterization, we compared the quantity and quality of the prepared libraries and the resulting sequences, and concordance of the overall results. We also assessed the impact of its implementation on laboratory workflow. The MagicPrep NGS produced higher library concentrations with smaller sizes, and correspondingly, higher molarity. Quality metrics of the sequences, however, demonstrated no significant impact on the overall results, producing 100% concordance with the reference method. Importantly, workflow analysis showed 5 hours less hands-on time per run with more flexibility. This evaluation study indicates that performance of the MagicPrep NGS is comparable to the Nextera DNA Flex with the added benefit of improving workflow efficiency and reducing labor for performing routine clinical microbial whole-genome sequencing tests.

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来源期刊
CiteScore
8.10
自引率
2.40%
发文量
143
审稿时长
43 days
期刊介绍: The Journal of Molecular Diagnostics, the official publication of the Association for Molecular Pathology (AMP), co-owned by the American Society for Investigative Pathology (ASIP), seeks to publish high quality original papers on scientific advances in the translation and validation of molecular discoveries in medicine into the clinical diagnostic setting, and the description and application of technological advances in the field of molecular diagnostic medicine. The editors welcome for review articles that contain: novel discoveries or clinicopathologic correlations including studies in oncology, infectious diseases, inherited diseases, predisposition to disease, clinical informatics, or the description of polymorphisms linked to disease states or normal variations; the application of diagnostic methodologies in clinical trials; or the development of new or improved molecular methods which may be applied to diagnosis or monitoring of disease or disease predisposition.
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