FOOD SCIENCE ›› 2022, Vol. 43 ›› Issue (12): 325-328.doi: 10.7506/spkx1002-6630-20210304-060
• Safety Detection • Previous Articles
LIU Jianhui, ZHANG Xianzhou, ZHANG Yunzhe, LI Lanru, WANG Hongjing, GAO Jie, GENG Fengzhen, TAN Jianxin
Published:
Abstract: In this study, a method to detect Salmonella was developed using polymerase chain reaction (PCR) combined with rolling circle amplification (RCA) with a dumbbell ring-shaped template into which a G-quadruplex complementary sequence was inserted. This method was based on PCR-RCA double amplification and the fact that thioflavin T (ThT) can specifically bind to the G-quadruplex, enhancing the fluorescence signal. Under the optimized detection conditions, the logarithmic concentration of Salmonella genomic DNA (x) had a good linear relationship with the fluorescence signal intensity at 486 nm (y). The regression equation was y = 2.101x + 2.872 3 (R2 = 0.992 5) in a linear range of 17 fg/μL–1.7 ng/μL. The specificity of the method was evaluated, revealing that it was suitable for the detection of Salmonella with a detection limit of 4.28 CFU/mL for artificially contaminated milk samples. This method, which has several advantages such as strong specificity, high sensitivity and low detection limit, provides a new method for the rapid detection of foodborne pathogens.
Key words: G-quadruplet; polymerase chain reaction; rolling circle amplification; Salmonella; fluorescence signal detection
CLC Number:
TS207.4
LIU Jianhui, ZHANG Xianzhou, ZHANG Yunzhe, LI Lanru, WANG Hongjing, GAO Jie, GENG Fengzhen, TAN Jianxin. Detection of Salmonella by G-quadruplex-Based PCR-RCA Double Amplification[J]. FOOD SCIENCE, 2022, 43(12): 325-328.
0 / / Recommend
Add to citation manager EndNote|Ris|BibTeX
URL: https://www.spkx.net.cn/EN/10.7506/spkx1002-6630-20210304-060
https://www.spkx.net.cn/EN/Y2022/V43/I12/325