Abstract：
:Whole-genome sequencing is likely to become increasingly used by local clinical microbiology laboratories, where sequencing volume is low compared with national reference laboratories. Here, we describe a universal protocol for simultaneous DNA extraction and sequencing of numerous different bacterial species, allowing mixed species sequence runs to meet variable laboratory demand. We assembled test panels representing 20 clinically relevant bacterial species. The DNA extraction process used the QIAamp mini DNA kit, to which different combinations of reagents were added. Thereafter, a common protocol was used for library preparation and sequencing. The addition of lysostaphin, lysozyme or buffer ATL (a tissue lysis buffer) alone did not produce sufficient DNA for library preparation across the species tested. By contrast, lysozyme plus lysostaphin produced sufficient DNA across all 20 species. DNA from 15 of 20 species could be extracted from a 24-h culture plate, while the remainder required 48-72 h. The process demonstrated 100% reproducibility. Sequencing of the resulting DNA was used to recapitulate previous findings for species, outbreak detection, antimicrobial resistance gene detection and capsular type. This single protocol for simultaneous processing and sequencing of multiple bacterial species supports low volume and rapid turnaround time by local clinical microbiology laboratories.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Raven KE,Girgis ST,Akram A,Blane B,Leek D,Brown N,Peacock SJdoi
10.1038/s41598-020-80031-8subject
Has Abstractpub_date
2021-01-08 00:00:00pages
193issue
1issn
2045-2322pii
10.1038/s41598-020-80031-8journal_volume
11pub_type
杂志文章abstract：:After nearly a century of vaccination and six decades of drug therapy, tuberculosis (TB) kills more people annually than any other infectious disease. Substantial challenges to disease eradication remain among vulnerable and underserved populations. The Guarani-Kaiowá people are an indigenous population in Paraguay an...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-020-79621-3
更新日期：2021-01-13 00:00:00
abstract：:The transcriptional regulator JDP2 (Jun dimerization protein 2) has been identified as a prognostic marker for patients to develop heart failure after myocardial infarction. We now performed in vivo studies on JDP2-overexpressing mice, to clarify the impact of JDP2 on heart failure progression. Therefore, during birth...
journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-018-26052-w
更新日期：2018-05-16 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-018-26170-5
更新日期：2018-05-21 00:00:00
abstract：:The human brain has evolved specialised mechanisms to enable the rapid detection of threat cues, including emotional face expressions (e.g., fear and anger). However, contextual cues - such as gaze direction - influence the ability to recognise emotional expressions. For instance, anger paired with direct gaze, and fe...
journal_title：Scientific reports
pub_type： 杂志文章
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journal_title：Scientific reports
pub_type： 杂志文章
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journal_title：Scientific reports
pub_type： 杂志文章
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journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/srep24459
更新日期：2016-04-14 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
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journal_title：Scientific reports
pub_type： 杂志文章
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journal_title：Scientific reports
pub_type： 杂志文章
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更新日期：2015-06-05 00:00:00
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pub_type： 杂志文章
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pub_type： 杂志文章
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更新日期：2017-08-17 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/srep44595
更新日期：2017-03-15 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
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更新日期：2017-08-07 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/srep01375
更新日期：2013-01-01 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-018-30643-y
更新日期：2018-08-13 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
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更新日期：2018-06-22 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/s41598-020-80477-w
更新日期：2021-01-12 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
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journal_title：Scientific reports
pub_type： 杂志文章
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journal_title：Scientific reports
pub_type： 杂志文章
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journal_title：Scientific reports
pub_type： 杂志文章
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journal_title：Scientific reports
pub_type： 杂志文章
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更新日期：2017-02-16 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
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更新日期：2018-03-02 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
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更新日期：2017-07-14 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
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更新日期：2020-07-20 00:00:00
abstract：:An amendment to this paper has been published and can be accessed via a link at the top of the paper. ...
journal_title：Scientific reports
pub_type： 已发布勘误
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更新日期：2020-07-29 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
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journal_title：Scientific reports
pub_type： 杂志文章
doi：10.1038/srep30110
更新日期：2016-07-22 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
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更新日期：2019-12-12 00:00:00
