Microwave-accelerated method for ultra-rapid extraction of Neisseria gonorrhoeae DNA for downstream detection.
Nucleic 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.
Institute of Fluorescence and Department of Chemistry and Biochemistry, University of Maryland Baltimore County, Baltimore, MD 21202, USA.
Johan H
JH
Melendez
Institute of Fluorescence and Department of Chemistry and Biochemistry, University of Maryland Baltimore County, Baltimore, MD 21202, USA.
Tonya M
TM
Santaus
Institute of Fluorescence and Department of Chemistry and Biochemistry, University of Maryland Baltimore County, Baltimore, MD 21202, USA.
Gregory
G
Brinsley
Institute of Fluorescence and Department of Chemistry and Biochemistry, University of Maryland Baltimore County, Baltimore, MD 21202, USA.
Daniel
D
Kiang
Institute of Fluorescence and Department of Chemistry and Biochemistry, University of Maryland Baltimore County, Baltimore, MD 21202, USA.
Buddha
B
Mali
The Johns Hopkins Medical Institutions, Baltimore, MD 21205, USA.
Justin
J
Hardick
The Johns Hopkins Medical Institutions, Baltimore, MD 21205, USA.
Charlotte A
CA
Gaydos
Institute of Fluorescence and Department of Chemistry and Biochemistry, University of Maryland Baltimore County, Baltimore, MD 21202, USA. Electronic address: geddes@umbc.edu.
Chris D
CD
Geddes
D004269:DNA, Bacterial / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D008872:Microwaves; D009344:Neisseria gonorrhoeae / Q000737:chemistry*
D004269:DNA, Bacterial
DNA extraction; DNA fragmentation; Gonorrhea; Microwave-accelerated metal-enhanced fluorescence; Microwaves; Neisseria gonorrhoeae