1. Recombinase polymerase amplification assays for the identification of pork and horsemeat.
- Author
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Kissenkötter J, Böhlken-Fascher S, Forrest MS, Piepenburg O, Czerny CP, and Abd El Wahed A
- Subjects
- Animals, DNA metabolism, Mitochondria metabolism, Mitochondrial Proton-Translocating ATPases genetics, NADH Dehydrogenase genetics, Point-of-Care Systems, Recombinases metabolism, DNA analysis, Horses genetics, Meat analysis, Nucleic Acid Amplification Techniques methods, Swine genetics
- Abstract
Detection of animal species in meat product is crucial to prevent adulterated and unnecessary contamination during processing. Gold standard is the real-time PCR assays, which can be conducted at highly equipped laboratories. Toward the development of point-of-need test, two rapid molecular assays based on recombinase polymerase amplification (RPA) for the detection of pork and horse DNA were established. Target genes are the porcine mitochondrial ND2 and equine ATP 6-8 genes. The pork and horse_RPA assays detected 16 and one DNA molecules/µl in eleven to six minutes, respectively. The myoglobin in the meat did not influence the assays performances, while the presence of high background-DNA induced a one log decrease in the sensitivity. Both assays are highly specific and identify down to 0.1% of their target DNA in meat mixtures. Both RPA assays could be used on-site as a rapid and mobile detection system to determine contamination of meat products., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Jonas Kissenkötter, Susanne Böhlken-Fascher, Claus-Peter CzernyandAhmed Abd El Waheddeclare no conflict ofinterest.Matthew S. Forrest and Olaf Piepenburg were employees at TwistDx Ltd. RPA technology is subject to background IP protection., (Copyright © 2020 Elsevier Ltd. All rights reserved.)
- Published
- 2020
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