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| Content Provider | frontiers |
|---|---|
| Author | Wang, Yi Li, Hui Wang, Yan Zhang, Lu Xu, Jianguo Ye, Changyun |
| Abstract | The report describes a simple, sensitive and nearly instrument-free assay for visual and multiplex detection of Enterococcus faecalis and Staphylococcus aureus based on multiple loop-mediated isothermal amplification (mLAMP) and lateral flow biosensor (LFB). Detection and differentiation of the Ef0027 gene (E. faecalis-specific gene) and nuc gene (S. aureus-specific gene) were determined using fluorescein (FITC)-and digoxin-modified primers in the mLAMP process. In the presence of biotin- and FITC-/digoxin-modified primers, the mLAMP yielded numerous biotin- and FITC-/digoxin-attached duplex products, which were detected by LFB through biotin/streptavidin interaction (biotin on the duplex and streptavidin on the gold nanoparticle) and immunoreactions (FITC/digoxin on the duplex and anti-FITC/digoxin on the LFB test line). The accumulation of gold nanoparticles generated a characteristic red line, enabling visual and multiplex detection of target pathogens without instrumentation. The analytical sensitivity, specificity and feasibility of LAMP-LFB technique were successfully examined in pure culture and blood samples. The entire procedure, including specimen (such blood samples) processing (30 min), isothermal reaction (40 min) and result reporting (within 2 min), could be completed within 75 min. Thus, this assay offers a simple, rapid, sensitive and specific test for multiplex detection of E. faecalis and S. aureus strains. Furthermore, the LAMP-LFB strategy is a universal technique, which can be extended to detect various target sequences by re-designing the specific LAMP primers. |
| ISSN | 1664302X |
| DOI | 10.3389/fmicb.2017.00192 |
| Volume Number | 8 |
| Journal | Frontiers in Microbiology |
| Language | English |
| Publisher Date | 2017-02-10 |
| Access Restriction | Open |
| Subject Keyword | Loop-mediated isothermal amplification E. faecalis LAMP-LFB S. aureus Limit of detecton Lateral flow biosensor |
| Content Type | Text |
| Resource Type | Article |
| Subject | Microbiology Microbiology (medical) |
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