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Development of real-time PCR methods for cocoa authentication in processed cocoa-derived products
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NSTL
Elsevier
The applicability of real-time PCR to highly processed cocoa-derived products for authentication purposes depends on the possibility of extracting high quality DNA and developing efficient PCR tests. This study compares three methods for DNA extraction from cocoa-derived products and chocolate. The best method providing suitable DNA for PCR amplification was the classical CTAB method. Seven real-time PCR tests for cocoa detection were designed based on low (nuclear) and high-copy (ribosomal and chloroplast) DNA targets. Two methods, one based on a nuclear target (vicilin-li PCR test) and a second one based on chloroplast DNA (lipids PCR test), successfully passed the performance criteria considering the specificity, sensitivity, efficiency of amplification, robustness and applicability in processed cocoa-derived products and chocolate. Therefore, this methodology is useful for cocoa authentication purposes and constitutes a first step in the direction of a future distinction between fine and bulk genotypes with an easy, fast and cost-effective methodology.
NSTL
Elsevier
