A Low-Cost Closed-Tube Method for Detection of Adulteration in Ground Meat

The increased public concern about adulteration and mislabelling of meat products has fueled a need for the development of fast, reliable, and cost-effective species detection methods. Hence, we developed a high resolution melt analysis (HRMA) for the discrimination of meat species commonly used in the meat industry and their possible adulterants. A universal primer pair spanning a ⁓238 bp fragment of mitochondrial 16S rRNA gene was designed to perform HRMA. The developed workflow was validated on standard meat samples, ternary meat mixtures of eight different species, and 56 commercial meat samples. Distinct melting profiles were generated for each species, several ternary meat mixtures, and commercial food samples. The assay was sufficiently sensitive to detect 0.003 ng/µl of DNA of every targeted species. In short, the developed HRM analysis is a rapid, cost-effective, and efficient species discrimination system for confirmation of meat origin.</p

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Overview of the sequence counts at different stages of the analysis.

a<p>Direct and reverse sequence reads corresponding to a single DNA molecule were aligned and merged, producing what we called a “properly assembled sequence”.</p>b<p>Strictly identical sequences correspond to “unique sequence”.</p

Location of Tost Mountain study site in South Gobi, Mongolia.

<p>Location of Tost Mountain study site in South Gobi, Mongolia.</p

A comparison of frequency of occurrence (percent) of various prey items in the feces of snow leopard from various regions of its range.

<p>A comparison of frequency of occurrence (percent) of various prey items in the feces of snow leopard from various regions of its range.</p

Unique sequences generated initially by NGS sequencing

contains files with unique sequences generated by next generation sequencin

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Contains file having data after filtration. The final result file for common leopard diet in Ayubia National Park, Pakista