Visual Detection of Salmonella by G-Quadruplex-Generating Polymerase Chain Reaction

被引：0
作者：
Liu J. [1 ]
Geng F. [2 ]
Zhang X. [1 ]
Gao H. [1 ]
Li C. [1 ]
Gao J. [1 ]
Tan J. [1 ]
机构：
[1] College of Food Science and Technology, Hebei Agricultural University, Baoding
[2] Affiliated Hospital of Hebei University, Baoding
来源：
Shipin Kexue/Food Science | 2021年 / 42卷 / 22期
关键词：
G-quadruplex; Polymerase chain reaction; Salmonella enteriditis; Visual detection;
D O I：
10.7506/spkx1002-6630-20200918-248
中图分类号：
学科分类号：
摘要：
In order to establish a visual detection method for Salmonella using G-guadruplex-generating polymerase chain reaction (PCR), the upstream and downstream primers containing G-quadruplex complementary sequences at the 5' end were designed according to the Salmonella-specific gene invH, which served as the target gene.Double-stranded DNA contained a large number of G-quadruplexes was obtained after specific amplification of the target sequences by PCR, and bound to hemin after being denaturated to generate DNAzyme with peroxidase-like activity.DNAzyme was able to catalyze the reaction between H2O2 and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, resulting in color change from colorless to green and thus allowing visual detection of Salmonella. Under the optimized detection conditions,there was a good linear relationship between the logarithm of Salmonella genome concentration (y) and absorbance at 421 nm (x), which was fitted to the equation: y=0.129 9x + 0.217 9 (R2=0.994 5) with a linear range of 0.07-771.6 ng/μL,and the sensitivity was 0.07 ng/μL.The specificity analysis showed that this method was suitable for the detection of Salmonella, and could be successfully applied to artificially contaminated milk samples, with a detection limit of 1.2 ×102 CFU/mL. By using this method, 30 commercial samples were tested and the results were consistent with those obtained by the national standard detection method. This study provides a new strategy for the detection of foodborne pathogens. © 2021, China Food Publishing Company. All right reserved.
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页码：331 / 338
页数：7
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