Development of a capture sequencing assay for enhanced detection and genotyping of tick-borne pathogens.
Development of a capture sequencing assay for enhanced detection and genotyping of tick-borne pathogens.
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开发捕获测序测定法，加强蜱媒病原体的检测和基因分型。
DOI：
10.1038/s41598-021-91956-z
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发表时间：
2021-06-11
影响因子：
4.6
通讯作者：
Tokarz R
中科院分区：
文献类型：
--
作者：
Jain K;Tagliafierro T;Marques A;Sanchez-Vicente S;Gokden A;Fallon B;Mishra N;Briese T;Kapoor V;Sameroff S;Guo C;Marcos LA;Hu L;Lipkin WI;Tokarz R
灵敏度不足一直是实施高通量测序研究蜱传病原体的主要限制。在这里，我们描述了TBDCapSeq的发展，这是一种测序检测方法，使用杂交捕获探针，覆盖了美国发现的11种最常见的蜱传病原体的完整基因组。探针用于基于溶液的病原体核酸捕获和富集，然后进行高通量测序。我们评估了TBDCapSeq监测包括人全血、小鼠组织和现场收集的蜱的样品的性能。对于伯氏疏螺旋体和田鼠巴氏疏螺旋体，TBDCapSeq的灵敏度是相当的，并且偶尔超过试剂特异性定量PCR的性能，并且当与标准无偏测序相比时，导致病原体读数增加25至> 10，000倍。TBDCapSeq还能够直接在脊椎动物和蜱宿主中进行基因组分析。TBDCapSeq的实施可能会对蜱传病原体的研究产生重大影响，通过改进检测和促进基因组研究，这在以前是无法用标准测序方法实现的。
Inadequate sensitivity has been the primary limitation for implementing high-throughput sequencing for studies of tick-borne agents. Here we describe the development of TBDCapSeq, a sequencing assay that uses hybridization capture probes that cover the complete genomes of the eleven most common tick-borne agents found in the United States. The probes are used for solution-based capture and enrichment of pathogen nucleic acid followed by high-throughput sequencing. We evaluated the performance of TBDCapSeq to surveil samples that included human whole blood, mouse tissues, and field-collected ticks. For Borrelia burgdorferi and Babesia microti, the sensitivity of TBDCapSeq was comparable and occasionally exceeded the performance of agent-specific quantitative PCR and resulted in 25 to > 10,000-fold increase in pathogen reads when compared to standard unbiased sequencing. TBDCapSeq also enabled genome analyses directly within vertebrate and tick hosts. The implementation of TBDCapSeq could have major impact in studies of tick-borne pathogens by improving detection and facilitating genomic research that was previously unachievable with standard sequencing approaches.
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影响因子：
48
作者：
Langmead, Ben;Salzberg, Steven L.
通讯作者：
Salzberg, Steven L.
影响因子：
--
作者：
Jones, Kathryn L.;McHugh, Gail A.;Steere, Allen C.
通讯作者：
Steere, Allen C.
DOI：
10.1146/annurev-pathmechdis-012418-012751
发表时间：
2019-01-24
期刊：
Annual review of pathology
影响因子：
--
作者：
Gu W;Miller S;Chiu CY
通讯作者：
Chiu CY
影响因子：
11.8
作者：
Cerar, Tjasa;Strle, Franc;Strle, Klemen
通讯作者：
Strle, Klemen
影响因子：
4.4
作者：
Marques, Adriana R.
通讯作者：
Marques, Adriana R.