Fast dissemination of the mobilized colistin resistance () gene in causes a huge threat to the treatment of severe infection. In the current report, a multiple cross displacement amplification (MCDA) coupled with the detection of amplified products by gold nanoparticles-based lateral flow biosensor (LFB) assay (MCDA-LFB) was established to identify the gene with simpleness, rapidity, specificity, and sensitivity. The MCDA-LFB assay was performed at a isothermal temperature (63°C) for only 30 min during the amplification stage, and the reaction products were directly identified by using LFB which obtained the result within 2 min. The entire process of experiments, from templates extraction to result judging, was accomplished in <60 min. For the analytical specificity of this method, all of the 16 -producing strains were positive, and all of the non- isolates produced the negative results. The sensitivity of -MCDA-LFB assay was as little as 600 fg of plasmid DNA per reaction in pure culture, and approximately 4.5 × 10 CFU/mL (~4.5 CFU/reaction) in spiked fecal samples. Therefore, this technique established in the present study is suitable for the surveillance of gene in clinic and livestock industry.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6610462 | PMC |
http://dx.doi.org/10.3389/fcimb.2019.00226 | DOI Listing |
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