Microwave-accelerated method for ultra-rapid extraction of Neisseria gonorrhoeae DNA for downstream detection

dc.contributor.authorMelendez, Johan
dc.contributor.authorSantaus, Tonya
dc.contributor.authorBrinsley, Gregory
dc.contributor.authorKiang, Daniel
dc.contributor.authorMali, Buddha
dc.contributor.authorHardick, Justin
dc.contributor.authorGaydos, Charlotte A.
dc.contributor.authorGeddes, Chris
dc.date.accessioned2024-10-01T18:05:28Z
dc.date.available2024-10-01T18:05:28Z
dc.date.issued2016-10-01
dc.description.abstractNucleic acid-based detection of gonorrhea infections typically require a two-step process involving isolation of the nucleic acid, followed by detection of the genomic target often involving polymerase chain reaction (PCR)-based approaches. In an effort to improve on current detection approaches, we have developed a unique two-step microwave-accelerated approach for rapid extraction and detection of Neisseria gonorrhoeae (gonorrhea, GC) DNA. Our approach is based on the use of highly focused microwave radiation to rapidly lyse bacterial cells, release, and subsequently fragment microbial DNA. The DNA target is then detected by a process known as microwave-accelerated metal-enhanced fluorescence (MAMEF), an ultra-sensitive direct DNA detection analytical technique. In the current study, we show that highly focused microwaves at 2.45 GHz, using 12.3-mm gold film equilateral triangles, are able to rapidly lyse both bacteria cells and fragment DNA in a time- and microwave power-dependent manner. Detection of the extracted DNA can be performed by MAMEF, without the need for DNA amplification, in less than 10 min total time or by other PCR-based approaches. Collectively, the use of a microwave-accelerated method for the release and detection of DNA represents a significant step forward toward the development of a point-of-care (POC) platform for detection of gonorrhea infections.
dc.description.sponsorshipThis work was supported in whole or part by NIH/NIBIB 5U54EB007958-09, NIH/NIGMS T32GM066706, and TEDCO MII Phase 1:#0115-010_2 awarded to CDG
dc.description.urihttps://www.sciencedirect.com/science/article/pii/S0003269716301440
dc.format.extent19 pages
dc.genrejouranl articles
dc.genrepostprints
dc.identifierdoi:10.13016/m2xvlm-1zxg
dc.identifier.citationMelendez, Johan H., Tonya M. Santaus, Gregory Brinsley, Daniel Kiang, Buddha Mali, Justin Hardick, Charlotte A. Gaydos, and Chris D. Geddes. ‘Microwave-Accelerated Method for Ultra-Rapid Extraction of Neisseria Gonorrhoeae DNA for Downstream Detection’. Analytical Biochemistry 510 (1 October 2016): 33–40. https://doi.org/10.1016/j.ab.2016.06.017.
dc.identifier.urihttps://doi.org/10.1016/j.ab.2016.06.017
dc.identifier.urihttp://hdl.handle.net/11603/36576
dc.language.isoen_US
dc.publisherElsevier
dc.relation.isAvailableAtThe University of Maryland, Baltimore County (UMBC)
dc.relation.ispartofUMBC Institute of Fluorescence (IoF)
dc.relation.ispartofUMBC Faculty Collection
dc.relation.ispartofUMBC Chemistry & Biochemistry Department
dc.relation.ispartofUMBC Student Collection
dc.rightshttps://creativecommons.org/licenses/by-nc-nd/4.0/
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subjectMicrowaves
dc.subjectDNA fragmentation
dc.subjectGonorrhea
dc.subjectDNA extraction
dc.subjectMicrowave-accelerated metal-enhanced fluorescence
dc.titleMicrowave-accelerated method for ultra-rapid extraction of Neisseria gonorrhoeae DNA for downstream detection
dc.typeText
dcterms.creatorhttps://orcid.org/0000-0002-8643-2343
dcterms.creatorhttps://orcid.org/0000-0002-9110-6374

Files

Original bundle

Now showing 1 - 1 of 1
Loading...
Thumbnail Image
Name:
nihms828365.pdf
Size:
960.08 KB
Format:
Adobe Portable Document Format