Fabrication of gold/silver nanodimer SERS probes for the simultaneous detection of Salmonella typhimurium and Staphylococcus aureus.

Salmonella typhimurium (S. typhimurium) and Staphylococcus aureus (S. aureus) are the two most important foodborne pathogens which can easily cause disease infections. Here, the aptamer-facilitated gold/silver nanodimer SERS probes were built for the simultaneous detection of the two bacteria with the help of magnetic separation enrichment. First, two nanodimer SERS signal probes and two magnetic capture probes each connected with the specific aptamer were fabricated. The distance between gold and silver nanoparticles in the dimer can amplify the Raman signal (Cy3 and Rox) at the junction but modified in the aptamer sequence. Then, after the addition of S. typhimurium and S. aureus, the sandwich-like composite structures "SERS signal probes-target-magnetic capture probes" formed because of the high affinity between aptamer sequences and their target bacteria. Under the optimal experimental conditions, the linear correlations between Raman intensity and the logarithm of the concentration of bacteria were y = 876.95x-67.84 (R2 = 0.9865) for S. typhimurium and y = 1280.43x-1752.6 (R2 = 0.9883) for S. aureus. The SERS detection showed the nanodimer probe had high selectivity. Besides, the recovery experiment in milk sample indicated good accuracy compared with the traditional plate counting method.

Drug Resistance and Integron Genes in Escherichia coli Isolated from Urinary Tract Infection.

Escherichia coli (E. coli) is a major cause of urinary tract infections. Treatment of these infections with antibiotics is often not effective due to the acquisition of drug-resistance genes by the bacteria. This process is mediated by integrons which belong to bacterial mobile genetic elements. Therefore, the present study addressed the issue of the relation between antibiotic resistance and integron genes in E. coli isolated from patients affected by urinary tract infection. Multiplex PCR assay employed to detect the E. coli integrase gene demonstrated that out of 49 bacterial strains, 26 were carrying class 1 integron and there was no case of bacteria harboring class 2 or class 3 integrons. Correlation analysis documented that E. coli strains harboring class 1 integron exhibited higher resistance towards tobramycin. The variable region gene cassette contained combinations of four genes responsible for antibiotic resistance: dfr17, aadA2, aadA5, and aac(6')-Ib-cr, of which the latter conferred tobramycin resistance. Together, the collected data underscore the need for identification and analysis of integrons in E. coli-induced urinary infections.

Aptamer based SERS detection of Salmonella typhimurium using DNA-assembled gold nanodimers.

The authors describe a surface-enhanced Raman scattering (SERS) based aptasensor for Salmonella typhimurium (S. typhimurium). Gold nanoparticles (AuNPs; 35 nm i.d.) were functionalized with the aptamer (ssDNA 1) and used as the capture probe, while smaller (15 nm) AuNPs were modified with a Cy3-labeled complementary sequence (ssDNA 2) and used as the signalling probe. The asymmetric gold nanodimers (AuNDs) were assemblied with the Raman signal probe and the capture probe via hybridization of the complementary ssDNAs. The gap between two nanoparticles is a "hot spot" in which the Raman reporter Cy3 is localized. It experiences a strong enhancement of the electromagnetic field around the particle. After addition of S. typhimurium, it will be bound by the aptamer which therefore is partially dehybridized from its complementary sequence. Hence, Raman intensity drops. Under the optimal experimental conditions, the SERS signal at 1203 cm-1 increases linearly with the logarithm of the number of colonies in the 102 to 107 cfu·mL-1 concentration range, and the limit of detection is 35 cfu·mL-1. The method can be performed within 1 h and was successfully applied to the analysis of spiked milk samples and performed very well and with high specificity. Graphical abstract DNA-assembled asymmetric gold nanodimers (AuNDs) were synthesized and appllied in a SERS-based aptasensor for S. typhimurium. Capture probe was preferentially combined with S. typhimurium and the structure of the AuNDs was destroyed. The "hot spot" vanished partly, this resulting in the decreased Raman intensity of Cy3.