Microwave-Accelerated Metal-Enhanced Fluorescence (MAMEF): A Rapid, < 10 Copy Number Detection Platform
In this chapter, we review a three-piece assay called microwave-accelerated metal-enhanced fluorescence (MAMEF). This method is rapid and efficient for detecting various proteins and DNA/RNA fragments from a range of bacteria. Currently, bacterial detection and identification are underpinned by robust instrumentation and cell cultures. Molecular detection strategies are available to detect DNA/RNA and proteins in time periods that range from hours to days. These strategies use various reagents, costly instrumentation, and can be time-consuming.
The MAMEF assay eliminates the need to amplify DNA and culturing methods for detection and has been developed by our research group over several years for the detection of various bacteria. The assay is based on the principles of metal-enhanced fluorescence and has undergone extensive experimental testing and clinical validations. In this chapter, we subsequently review the overarching principles behind MAMEF, the development of protein assays for anthrax and myoglobin, and DNA hybridization assays for Salmonella, Chlamydia, and Gonorrhea. Microwave-accelerated metal-enhanced fluorescence offers the significant benefit of rapid protein and DNA/RNA detection without the need for thermo-cycling-based amplification and culturing methods, which are widely used today, despite their significant run times.
KeywordsMicrowave-accelerated metal-enhanced fluorescence DNA detection Protein detection Rapid detection Low-copy number detection
- 1.Gaydos CA, Cartwright CP, Colaninno P, Welsch J, Holden J, Ho SY, Webb EM, Anderson C, Bertuzis R, Zhang L, Miller T, Leckie G, Abravaya K, Robinson J (2010) Performance of the Abbott RealTime CT/NG for detection of Chlamydia trachomatis and Neisseria gonorrhoeae. J Clin Microbiol 48(9):3236–3243CrossRefGoogle Scholar
- 2.Gaydos CA, Quinn TC, Willis D, Weissfeld A, Hook EW, Martin DH, Ferrero DV, Schachter J (2003) Performance of the APTIMA Combo 2 assay for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in female urine and endocervical swab specimens. J Clin Microbiol 41(1):304–309CrossRefGoogle Scholar
- 3.Van Der Pol B, Liesenfeld O, Williams JA, Taylor SN, Lillis RA, Body BA, Nye M, Eisenhut C, Hook EW III (2012) Performance of the Cobas CT/NG test compared to the Aptima AC2 and Viper CTQ/GCQ assays for detection of chlamydia trachomatis and Neisseria gonorrhoeae. J Clin Microbiol 50:2244–2249CrossRefGoogle Scholar
- 11.Aslan K, Holley P, Geddes CD (2006) Research paper: microwave-accelerated metal-enhanced fluorescence (MAMEF) with silver colloids in 96-well plates: application to ultra fast and sensitive immunoassays, high throughput screening and drug discovery. J Immunol Methods 312:137–147CrossRefGoogle Scholar
- 14.Newby LK, Storrow AB, Gibler WB, Garvey JL, Tucker JF, Kaplan AL, Schreiber DH, Tuttle RH, McNulty SE, Ohman EM (2001) Bedside multimarker testing for risk stratification in chest pain units: the chest pain evaluation by creatine kinase-MB, myoglobin, and troponin I (CHECKMATE) study. Circulation 103(14):1832–1837CrossRefGoogle Scholar
- 27.Levy H, Diallo S, Tennant SM, Livio S, Sow SO, Tapia M, Fields PI, Mikoleit M, Tamboura B, Kotloff KL, Lagos R, Nataro JP, Galen JE, Levine MM (2008) PCR method to identify Salmonella enterica serovars Typhi, Paratyphi A, and Paratyphi B among Salmonella isolates from the blood of patients with clinical enteric fever. J Clin Microbiol 46(5):1861–1866CrossRefGoogle Scholar
- 33.Tennant SM, Diallo S, Levy H, Livio S, Sow SO, Tapia M, Fields PI, Mikoleit M, Tamboura B, Kotloff KL, Nataro JP, Galen JE, Levine MM (2010) Identification by PCR of non-typhoidal Salmonella enterica serovars associated with invasive infections among febrile patients in Mali. PLoS Negl Trop Dis 4(3):1–9CrossRefGoogle Scholar