GenSeizer: a Multiplex PCR-Based Targeted Gene Sequencing Platform for Rapid and Accurate Identification of Major Mycobacterium Species.
GenSeizer: a Multiplex PCR-Based Targeted Gene Sequencing Platform for Rapid and Accurate Identification of Major Mycobacterium Species.
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GenSeizer：基于多重 PCR 的靶向基因测序平台，可快速准确地鉴定主要分枝杆菌物种。
DOI：
10.1128/jcm.00584-20
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发表时间：
2021-01-21
影响因子：
9.4
通讯作者：
Chu H
中科院分区：
文献类型：
--
作者：
Li B;Xu L;Guo Q;Chen J;Zhang Y;Huang W;Zhang Z;Han L;Xu X;Chu H
结核分枝杆菌和非结核分枝杆菌（NTM）感染常常表现出相似的临床症状。及时有效的治疗依赖于对菌种和耐药基因型的快速准确鉴定。
在这项研究中，开发了一种新的平台（GenSeizer），它结合了对大数据集的生物信息学分析和基于多重PCR的靶向基因测序，用于鉴定导致肺部以及肺外人类疾病的10种主要分枝杆菌菌种。同时检测脓肿分枝杆菌中某些erm(41)和rrl耐药基因型也是可行的。该平台具有特异性和敏感性，在参考菌株之间无交叉反应，检测限为5个DNA拷贝或50 CFU分枝杆菌/毫升。在一次盲法比较中，GenSeizer和多基因测序在鉴定88株临床分枝杆菌分离株的能力上显示出100%的一致性。通过对30株脓肿分枝杆菌菌株进行全基因组测序所确认的耐药基因型，也被GenSeizer 100%正确鉴定。这些结果表明GenSeizer是一种用于检测主要致病性分枝杆菌菌种的高效、可靠的平台。
Mycobacterium tuberculosis and nontuberculous mycobacterium (NTM) infections often exhibit similar clinical symptoms. Timely and effective treatment relies on the rapid and accurate identification of species and resistance genotypes. ABSTRACT Mycobacterium tuberculosis and nontuberculous mycobacterium (NTM) infections often exhibit similar clinical symptoms. Timely and effective treatment relies on the rapid and accurate identification of species and resistance genotypes. In this study, a new platform (GenSeizer), which combines bioinformatics analysis of a large data set and multiplex PCR-based targeted gene sequencing, was developed to identify 10 major Mycobacterium species that cause pulmonary, as well as extrapulmonary, human diseases. The simultaneous detection of certain erm(41) and rrl resistance genotypes in M. abscessus was also feasible. This platform was specific and sensitive and exhibited no cross-reactivity among reference strains and a detection limit of 5 DNA copies or 50 CFU Mycobacterium/ml. In a blind comparison, GenSeizer and multigene sequencing showed 100% agreement in the ability to identify 88 clinical Mycobacterium isolates. The resistance genotypes, confirmed by whole-genome sequencing of 30 M. abscessus strains, were also correctly identified by GenSeizer 100% of the time. These results indicate that GenSeizer is an efficient, reliable platform for detecting major pathogenic Mycobacterium species.