TaqMan qPCR for detection and quantification of mitochondrial DNA from toxic pufferfish species

• The designed TaqMan primer and probe set can detect at least 4 pufferfish genera.
• The detection limit is at as low as 1.75 pg of gDNA.
• Developed assay can specifically detect and quantify pufferfish gDNA within 1 h.

Outbreaks of pufferfish food poisoning have been reported worldwide, most were from unsuitable food preparation and adulteration. In order to rapidly detect pufferfish adulterant in processed foods, we developed and proposed the use of a TaqMan probe-based quantitative PCR. The designed detection oligos targeted a unique region in 16S rDNA of toxic marine pufferfish in Tetraodontidae Family and gave a positive signal at ≥1.75 pg of genomic DNA. Non-target DNA samples from other fish, chicken, and beef were negative.