In this study, we developed the genetically modified organism detection method by using the combination of rolling circle amplification (RCA) and surface-enhanced Raman spectroscopy (SERS) and compared the developed RCA-SERS assay with traditional-SERS assay. As part of the study, gold surfaces and gold nanorods were used to form sandwich structure. Gold nanorods were labeled with 5,5’-Dithiobis(2- Nitrobenzoic acid) (DTNB) to enhance SERS intensity. Quantification of the target concentration was performed via SERS spectra of DTNB on the nanorods. SERS spectra of target molecules were enhanced through the RCA reaction and the detection limit was found to be 6.3 fM. The sensitivity of developed RCA-SERS assay was compared with traditional SERS assay and the detection limit was found to be 0.1 pM. For the developed RCA-SERS assay, the control studies were performed using the 35S promoter of Bt-176 maize gene and nonsense sequence. It was found out that the developed RCA-SERS sandwich assay method is quite sensitive, selective and specific for target sequences in model and real systems.

Comparison between traditional-SERS and RCA-SERS assays for 35S promoter gene detection

251st National Meeting (2016)

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