Ultrasensitive SERS detection of Bacillus thuringiensis special gene based on Au@Ag NRs and magnetic beads

Highly sensitive and selective detection of specific DNA sequences is of great importance in clinical diagnosis, environmental and food monitoring, but it still remains challenges to develop a facile method for real sample detection in aqueous solution. Here, a simple and recyclable surface enhanced Raman scattering (SERS) sensor was constructed for Bacillus thuringiensis (Bt) special gene fragment detection by Fe₃O₄ magnetic beads (MBs) and Au-Ag core-shell nanorods (Au@Ag NRs). A hairpin DNA with sulfhydryl and biotin was attached to Au@Ag NRs as indicator, and MBs with streptavidin (SA) were acted as the capture probe. On the basis of the biotin-SA specific interaction, target sequences were first hybridized with the hairpin DNA and exposed the biotin. Subsequently, the Au@Ag NRs were captured by the streptavidin modified MBs, which reduced the suspended NRs and led to the change of Raman intensity. Under the optimal conditions, the SERS intensity revealed a good linearity with Bt transgene fragment ranging from 0.1pM to 1nM with a detection limit of 0.14pM (S/N=3). To demonstrate the specificity of the strategy, the single-base mismatch in DNA was discussed in the SERS assay. The results showed that the sensitivity and accuracy of the proposed method was acceptable in DNA detection, revealing a great potential in special gene detection.