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Implementation of antimicrobial peptides for sample preparation prior to nucleic acid amplification in point-of-care settings
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Number of Authors: 72017 (English)In: Expert Review of Molecular Diagnostics, ISSN 1473-7159, E-ISSN 1744-8352, Vol. 17, no 12, p. 1117-1125Article in journal (Refereed) Published
Abstract [en]

Background: A variety of sample preparation techniques are used prior to nucleic acid amplification. However, their efficiency is not always sufficient and nucleic acid purification remains the preferred method for template preparation. Purification is difficult and costly to apply in point-of-care (POC) settings and there is a strong need for more robust, rapid, and efficient biological sample preparation techniques in molecular diagnostics. Methods: Here, the authors applied antimicrobial peptides (AMPs) for urine sample preparation prior to isothermal loop-mediated amplification (LAMP). AMPs bind to many microorganisms such as bacteria, fungi, protozoa and viruses causing disruption of their membrane integrity and facilitate nucleic acid release. Results: The authors show that incubation of E. coli with antimicrobial peptide cecropin P1 for 5 min had a significant effect on the availability of template DNA compared with untreated or even heat treated samples resulting in up to six times increase of the amplification efficiency. Conclusion: These results show that AMPs treatment is a very efficient sample preparation technique that is suitable for application prior to nucleic acid amplification directly within biological samples. Furthermore, the entire process of AMPs treatment was performed at room temperature for 5 min thereby making it a good candidate for use in POC applications.

Place, publisher, year, edition, pages
2017. Vol. 17, no 12, p. 1117-1125
Keywords [en]
Antimicrobial peptides, LAMP, crude sample preparation, point-of-care, AMPs, loop mediated isothermal amplification
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy) Biological Sciences
Identifiers
URN: urn:nbn:se:su:diva-152670DOI: 10.1080/14737159.2017.1386557ISI: 000422752800008PubMedID: 28965426OAI: oai:DiVA.org:su-152670DiVA, id: diva2:1183730
Available from: 2018-02-19 Created: 2018-02-19 Last updated: 2018-02-19Bibliographically approved

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Langel, Ülo
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