Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8478710 | Molecular and Cellular Probes | 2018 | 17 Pages |
Abstract
Coupling propidium monoazide (PMA) with quantitative PCR (PMA-qPCR) has been successfully applied to specific detection and quantification of viable cells in various samples. The optimal PMA treatment condition is usually determined through qPCR. However, it is a tedious, time consuming and costly process including DNA extraction and qPCR. To overcome this problem, a flow cytometry-based (FCM-based) method was first proposed in this study to replace qPCR for screening of the optimal PMA treatment condition for Helicobacter pylori, since the pure culture treated with PMA was actually a single cell suspension with fluorescent dye. Results showed that the optimal PMA treatment condition (30â¯Î¼M of PMA and 8â¯min of exposure time) determined by the novel method was the same as that determined by the qPCR-based method, which demonstrate the feasibility of this approach. In addition, with the comparison of the qPCR-based method, the FCM-based method allows screening of the optimal PMA treatment condition become much more simple, rapid and economical.
Keywords
Related Topics
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Biochemistry, Genetics and Molecular Biology
Cell Biology
Authors
Zhiqing Huang, Jianwei Zheng, Chunmei Shi, Qiang Chen,