Abstract:
:New Delhi metallo-β-lactamase (NDM)-producing bacteria are considered potential global health threats. It is necessary to monitor NDM-1 and its variants in clinical isolates in order to understand the NDM-1 epidemic and the impact of its variants on β-lactam resistance. To reduce the lengthy time needed for cloning and expression of NDM-1 variants, a novel PCR-based in vitro protein expression (PCR-P) method was used to detect blaNDM-1 and its variants coding for carbapenemases with different activities (functional variants). The PCR-P method combined a long-fragment real-time quantitative PCR (LF-qPCR) with in vitro cell-free expression to convert the blaNDM-1 amplicons into NDM for carbapenemase assay. The method could screen for blaNDM-1 within 3 h with a detection limit of 5 copies and identify functional variants within 1 day. Using the PCR-P to analyze 5 recent blaNDM-1 variants, 2 functional variants, blaNDM-4 and blaNDM-5, were revealed. In the initial testing of 23 clinical isolates, the PCR-P assay correctly found 8 isolates containing blaNDM-1. This novel method provides the first integrated approach for rapidly detecting the full-length blaNDM-1 and revealing its functional variants in clinical isolates.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Huang L,Hu X,Zhou M,Yang Y,Qiao J,Wang D,Yu J,Cui Z,Zhang Z,Zhang XE,Wei Hdoi
10.1128/JCM.03363-13subject
Has Abstractpub_date
2014-06-01 00:00:00pages
1947-53issue
6eissn
0095-1137issn
1098-660Xpii
JCM.03363-13journal_volume
52pub_type
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
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更新日期:1987-10-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.35.2.511-514.1997
更新日期:1997-02-01 00:00:00
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pub_type: 临床试验,杂志文章
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更新日期:1995-04-01 00:00:00
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pub_type: 杂志文章
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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更新日期:1992-10-01 00:00:00
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更新日期:2000-07-01 00:00:00
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journal_title:Journal of clinical microbiology
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更新日期:1980-10-01 00:00:00