Mitochondrial DNA survival in leather /leather products and Taqman based
PCR assay explores the credibility of identification: An approach on
Ovis aries (Indian)
Abstract
Leather and leather products are the most traded commodities and play a
significant role in the world’s economy. The traditional tanning methods
and the high quality of Indian leathers receive global appreciation. The
existing leather manufacturing methods made a significant loss in
epidermal constituents which impact the commercial importance of the
animal species. Embossing techniques could potentially contribute to an
increase in fraudulent activities. Since the fate of genetic material in
finished leather is questionable, which poses a challenge to the
molecular approach for identification. In order to obviate the problem,
the present study aims to recover amplifiable mtDNA from leather
products and focus on developing probes and primers for RT-PCR, which
significantly enhance cross-specificity. Data mining and phylogenetic
analyses of mtDNA genes (12S rRNA, COI, and Cyt b) for Indian Ovis aries
skins exhibit the taxonomic positions and the intraspecies
relationships, thereby facilitating the probes desigining. The results
indicate that the maximum intraspecies genetic divergence observed
within 12S rRNA, followed by COI and Cyt b. Since the availability of
genetic sequences with reference to Cyt b is comparatively large (602),
primers and probes were designed for Cyt b. The RT-PCR analysis of
finished Ovis aries leather mtDNA samples yield a ct value in the range
of 22–23 cycles, even with a minimum quantity of template DNA as low as
0.3 ng. Results on the cross-specificity analysis are highly
encouraging. Hence, it is feasible to identify genuine Ovis aries
leather using mtDNA instilling confidence in the export of leather.