Recombinase polymerase amplification combined with lateral flow dipstick for equipment-free detection of Salmonella in shellfish.
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Abstract |
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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. |
Year of Publication |
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2018
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Journal |
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Bioprocess and biosystems engineering
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Date Published |
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2018
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ISSN Number |
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1615-7591
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URL |
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https://dx.doi.org/10.1007/s00449-018-1895-2
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DOI |
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10.1007/s00449-018-1895-2
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Short Title |
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Bioprocess Biosyst Eng
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