12400088893?profile=RESIZE_400xThis dissertation from Florida State University describes designing and validating a species-specific PCR-lateral flow assay for Atlantic white shrimp (Litopenaeus setiferus) utilizing a miniaturized and cost-effective PCR instrument.  The selectivity was tested against 68 shrimp, prawn, and fish samples from 14 seafood species. L. setiferus was simultaneously amplified by the multiplex assay to give three visual bands, which distinguished it from other species having either one or two bands on the lateral flow stick.

The researcher also developed a specific red snapper assay, validating a rhPCR lateral flow assay where the thermotolerant RNase H2 enzyme was included in the PCR reaction to activate the red snapper-specific rhPCR primer. Amplicons generated in the duplex rhPCR reaction were detected using dual target lateral flow strips. The standardized assay was validated with 108 barcoded fish samples from 16 finfish species. Samples identified as Lutjanus campechanus or L. purpureus by DNA barcoding formed three distinct bands, while other fish species formed only two bands on the lateral flow strips. A minimum of 0.37 ng/μL crude DNA was needed to obtain a visible band on the lateral flow dip stick.

Both assays showed 100% specificity and took 90–120 minutes for completion. The researcher concludes that this confirms the suitability of PCR and rhPCR-lateral flow assays as an economical on-site tool for species authentication in the seafood industry.

Photo by Michal Mrozek on Unsplash

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