Abstract
This proof-of-principle study describes the development of a rapid and easy-to-use DNA microarray assay for the authentication of giant tiger prawns and whiteleg shrimp. Following DNA extraction and conventional end-point PCR of a 16S rDNA segment, the PCR products are hybridised to species-specific oligonucleotide probes on DNA microarrays located at the bottom of centrifuge tubes (ArrayTubes) and the resulting signal patterns are compared to those of reference specimens. A total of 21 species-specific probes were designed and signal patterns were recorded for 47 crustacean specimens belonging to 16 species of seven families. A hierarchical clustering of the signal patterns demonstrated the specificity of the DNA microarray for the two target species. The DNA microarray can easily be expanded to other important crustaceans. As the complete assay can be performed within half a day and does not require taxonomic expertise, it represents a rapid and simple alternative to tedious DNA barcoding and could be used by crustacean trading companies as well as food control authorities for authentication of crustacean commodities.Graphical abstract
Highlights
Crustaceans such as shrimp and prawns are popular food commodities that are traded worldwide
The aim of the present study was to test whether two crustacean species, the giant tiger prawn (P. monodon) and the whiteleg shrimp (P. vannamei), can be detected using a DNA microarray assay followed by a hierarchical clustering comparison of probe signal patterns
This study has shown that DNA microarrays are a promising alternative to sequencing-based approaches for authentication of crustacean products with respect to the declared species
Summary
Crustaceans such as shrimp and prawns are popular food commodities that are traded worldwide. Since shrimp and prawn products are often exported and traded as processed products (e.g. peeled and cooked specimens), it is usually not possible to verify whether the purchased products are authentic in relation to the declared species without elaborate laboratory analyses. Species-level crustacean authentication methods are needed for both food control and food industry (or respective service laboratories) to verify and ensure proper labelling of traded crustacean products. The determined sequences have to be compared with DNA sequences in public databases such as GenBank [1] and BOLD [2] This DNA barcoding is time consuming and tedious, and requires considerable expertise in the taxon under study as well as the quality of the sequence data in the databases. Commissioning service laboratories to authenticate species authentication for self-monitoring analyses is quite expensive and the processing time usually takes several days
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