AN ADVANCED VISUAL QUALITATIVE AND EVA GREEN-BASED QUANTITATIVE ISOTHERMAL AMPLIFICATION METHOD TO DETECT LISTERIA MONOCYTOGENES

作者：Cheng, Nan; Xu, Yuancong; Yan, Xinghua; Shang, Ying; Zhu, Pengyu; Tian, Wenying; Liang, Zhihong; Xu, Wentao^*

来源：Journal of Food Safety, 2016, 36(2): 237-246.

DOI：10.1111/jfs.12236

• REAL-TIME PCR
• RAPID DETECTION
• DNA-POLYMERASE
• FOOD
• QUANTIFICATION
• IDENTIFICATION
• POINT
• ASSAY
• SPP.
• LAMP

The purpose of this study is to apply the visual qualitative (LFD-LAMP) and advanced quantitative loop-mediated isothermal amplification (EVA-LAMP) methods to detect Listeria monocytogenes, which is an important pathogenic foodborne bacterium. This research designed a new set of primers to amplify the hlyA gene of L.monocytogenes. The LFD-LAMP products were detected by lateral flow device (LFD) with the naked eyes, and the test results were consistent with agarose gel electrophoresis. Meanwhile, an advanced quantitative LAMP assay was established which depended on the EVA Green. The detection limit of the EVA-LAMP method was 10pg/L, and a good linear relationship was obtained. The subsequent analysis of contaminated raw food samples showed that the primers were able to identify L.monocytogenes in food. The LFD-LAMP and EVA-LAMP reactions can be used as sensitive, rapid and simple methods for the detection of L.monocytogenes, especially on field detection.

• 出版日期2016-5
• 单位中国农业大学