Using MinION Nanopore sequencing to simultaneously identify multiple, illegally imported wild meat samples in Brussels, Belgium
Authors
Ann Vanderheyden,
Sophie Gombeer,
Anne-Lise Chaber,
Kenny Meganck,
Nathalie Smitz,
Casimir Nebesse,
Prescott Musaba,
Steve Ngoy,
Thierry Backeljau,
Marc De Meyer,
Erik Verheyen,
Discipline
Biological sciences
Subject
Invertebrates
Audience
Scientific
Date
2022Metadata
Show full item recordDescription
The import of African wild meat into Europe (EU) is of concern because it contributes to the overexploitation of vulnerable and protected species in the regions of origin. In addition, it poses a health risk to humans, livestock, and local biodiversity in the destination countries through the possible transmission of exotic infectious diseases. Identifying wild meat at border controls, however, is challenging as it is often smoked or processed for consumption, so that no morphological features remain for species identification. Despite legislations forbidding the import of (wild) meat into the EU, several studies have demonstrated that large volumes of meat, mostly of mammalian origin, pass undetected through custom controls. BopCo (Barcoding facility for organisms and tissues of policy concern, https://bopco.myspecies.info/) uses DNA barcoding to identify species and tissues of policy concern. However, conventional DNA barcoding methods, via Sanger sequencing, can be expensive and time consuming when many samples need to be identified. Additionally, a DNA identification to species level often requires sequencing of multiple DNA markers. Using MinION Nanopore sequencing, BopCo developed a quick DNA barcoding protocol to identify wild meat samples based on a combination of four mitochondrial DNA markers (COI, Cytb, 16S and 12S). Using specific primer sets, all four markers are amplified in a multiplex PCR reaction. The MinION technology allows several samples to be pooled together during the library preparation, increasing the sequencing output. After demultiplexing the different samples, consensus sequences for the four markers are extracted by mapping the generated reads to a mitochondrial reference genome. This third generation sequencing technique allows for a rapid, efficient and reliable identification of a large number of samples in a cost-effective way.
Citation
Ann Vanderheyden,; Sophie Gombeer,; Anne-Lise Chaber,; Kenny Meganck,; Nathalie Smitz,; Casimir Nebesse,; Prescott Musaba,; Steve Ngoy,; Thierry Backeljau,; Marc De Meyer,; Erik Verheyen, (2022). Using MinION Nanopore sequencing to simultaneously identify multiple, illegally imported wild meat samples in Brussels, Belgium. , International Conference on DNA Barcoding and Biodiversity,Identifiers
Type
Conference
Peer-Review
No
Language
eng