Journal: Virology Journal
Article Title: A novel dengue virus detection method that couples DNAzyme and gold nanoparticle approaches
Figure Lengend Snippet: Assessment of DDZ-AuNP specificity. A) DENV-2 and CHIKV (1 × 106/mL each) were placed in a buffered solution containing 10 mM MgCl2, 1 × 105 DDZ-AuNP particles/mL, 1.5 M NaCl, and 0.5% (w/v) SDS. Eppendorf tubes containing these mixes were incubated at 37°C for 30 minutes and photographed. CHIKV = chikungunya virus. DENV-2 = dengue virus serotype 2, DDZ-M = anti-dengue virus DNAzyme, DDZin-M = inactive anti-dengue virus DNAzyme. B) Analysis of DENV detection by UV/Vis Spectrophotometry. Samples were assembled as was performed for in A, mixed, incubated at 37°C for 5 minutes, and spectrophotometric analysis was performed using the ND-1000 spectrophotometer. C) Detection of DENV by DDZ-M-AuNP in comparison to several other flaviviruses. The specificity of our DDZ-M-AuNP device to detect DENV and not other fellow flavivirus members YFV, JEV or ZV was determined as described earlier (see Figure 4 ). D) An alignment was performed on consensus sequences of each of the four DENV serotypes to determine the most optimal regions for the design of serotype specific DDZ-AuNP devices by determining the region of least conservation one serotype has with the other DENV serotypes. E) Colorimetric serotype-specific detection of DENV. Cell culture supernatants from C6/36 cells mock infected (Mock), or from cells infected with either DENV serotypes 1 through 4 or CHIKV (1 × 106/mL each) were placed in buffered solutions containing the necessary cofactors and AuNPs tethered with our all purpose DENV serotype specific DNAzyme, DDZ-M, or one of the serotype-specific DDZs (designated DDZ-1 through-4) designed to specifically target the corresponding DENV serotype (Table 1 ). Eppendorf tubes containing these mixes were incubated at 37°C for 5 minutes and photographed.
Article Snippet: Quantification of these oligonucleotides was performed with the ND-1000 spectrophotometer from NanoDrop (Wilmington, DE).
Techniques: Incubation, Spectrophotometry, Cell Culture, Infection