Recombinase polymerase amplification combined with lateral flow dipstick for equipment-free detection of Salmonella in shellfish

Salmonella is a major pathogen that causes acute foodborne outbreaks worldwide. Seafood, particularly shellfish, is a proven source of Salmonella spp. infection because many people prefer to eat it raw or lightly cooked. However, traditional identification methods are too time-consuming and complex...

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Veröffentlicht in:Bioprocess and biosystems engineering 2018-05, Vol.41 (5), p.603-611
Hauptverfasser: Gao, Weifang, Huang, Hailong, Zhu, Peng, Yan, Xiaojun, Fan, Jianzhong, Jiang, Jinpo, Xu, Jilin
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Sprache:eng
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Zusammenfassung:Salmonella is a major pathogen that causes acute foodborne outbreaks worldwide. Seafood, particularly shellfish, is a proven source of Salmonella spp. infection because many people prefer to eat it raw or lightly cooked. However, traditional identification methods are too time-consuming and complex to detect contamination of bacteria in the food chain in a timely manner, and few studies have aimed to identify Salmonella in shellfish early in the supply chain. We herein developed a method for rapid detection of Salmonella in shellfish based on the method of recombinase polymerase amplification (RPA) combined with lateral flow dipstick (LFD), which targets the invasion gene A ( invA ). The RPA-LFD was able to function at 30–45 °C, and at the temperature of 40 °C, it only took 8 min of amplification to reach the test threshold of amplicons. The established method had both a good specificity and a sensitivity of 100 fg DNA per reaction (20 µL). Regarding practical performance, RPA-LFD performed better than real-time PCR. Another advantage of RPA-LFD is that it was capable of being performed without expensive equipments. Thus, RPA-LFD has potential for further development as a detection kit for Salmonella in shellfish and other foods under field conditions.
ISSN:1615-7591
1615-7605
DOI:10.1007/s00449-018-1895-2