Construction of PCR-SERS Method for Detection of <i>Vibrio parahaemolyticus</i>
Antuo Hu,
Xiaoting Song,
Xiaojie Sun,
Zhaoxin Lu,
Xinmei Liu,
Xiaomei Bie,
Jun Yang
Affiliations
Antuo Hu
College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
Xiaoting Song
College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
Xiaojie Sun
Key Laboratory of Detection and Traceability Technology of Foodborne Pathogenic Bacteria for Jiangsu Province Market Regulation, Nanjing Institute for Food and Drug Control, Nanjing 211198, China
Zhaoxin Lu
College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
Xinmei Liu
Key Laboratory of Detection and Traceability Technology of Foodborne Pathogenic Bacteria for Jiangsu Province Market Regulation, Nanjing Institute for Food and Drug Control, Nanjing 211198, China
Xiaomei Bie
College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
Jun Yang
Key Laboratory of Detection and Traceability Technology of Foodborne Pathogenic Bacteria for Jiangsu Province Market Regulation, Nanjing Institute for Food and Drug Control, Nanjing 211198, China
A paper-based surface enhancement of a Raman scattering substrate consisting of silver-nanowires stacked on glass-fiber filter paper was prepared. At the same time, the DNA-embedding molecule Eva Green was introduced as a signaling molecule for surface-enhanced Raman scattering (SERS) detection. Polymerase chain reaction (PCR) was used to amplify target genes and the method was developed into a rapid molecular diagnostic system. The total detection time of the developed detection method was 40 min, including 30 min of PCR amplification and 10 min of SERS measurement. After 30 PCR cycles, bacterial DNA with an initial concentration of 20 fg/μL and a bacterial suspension with an initial concentration of 7.2 × 101 CFUs/mL could be detected. When the enrichment culture time was 4 h, target bacteria with an initial contamination inoculation volume of 1.5 CFUs/mL could be detected in artificially contaminated samples. The method is fast and highly sensitive, and has not been applied to the detection of V. parahaemolyticus.
