Molecular Authentication of Meats from Three Terrestrial Birds Based on Pcr-Rflp Analysis of the Mitochondrial 12S rRNA Gene

Abstract

In this study, a method utilizing PCR-restriction fragment length polymorphism (PCR-RFLP) of a mitochondrial gene was developed for the identification of chicken (Gallus gallus), quail (Coturnix coturnix), and common pigeon (Columba livia) meat. PCR products of ~440 bp were obtained from the 12S rRNA gene of these three birds using a pair of universal primers. The three terrestrial birds can be distinguished using one restriction endonuclease, Alu I, which was selected based on species-specific variations in the mt 12S rRNA gene sequence using 9 newly-obtained and 44 published chicken, quail and pigeon sequences. This method was also successfully used to identify commercial quail and pigeon meat products, which were found to be adulterated with chicken meat. Additionally, our method had relatively high sensitivity for detecting a meat mixture. Ten percent of chicken meat in the mixed quail and pigeon sample was detectable. This assay can be useful for the accurate identification of meats from terrestrial birds, avoiding mislabeling or fraudulent species substitution in meat products.

A systematic review of DNA‐based methods in authentication of game and less common meat species

“…The authors pointed out the ease with which the assay could be performed without expensive instrumentation. He et al (2018) confirmed the cheapness and simplicity of the restriction fragment length assay in their work authenticating pigeon and quail meat products. PCR-SFLP (satellite fragment length polymorphism), another variant of PCR-RFLP, was used in a study that confirmed the presence of other species in bison meat products (Scales et al, 2021).…”

“…He et al. (2018) confirmed the cheapness and simplicity of the restriction fragment length assay in their work authenticating pigeon and quail meat products.…”

Evaluation of High Resolution Melting (HRM) Analysis for Meat Species Identification of Raw and Cooked Meat

“…The PCR technique is widely used because of its high sensitivity, feasibility, and ease of use, and it is regarded as a reliable technique for detecting meat frauds [13]. Sensitive PCR assays as popular techniques for DNA detection can be used when primers target multicopy genes, including mitochondrial DNA (mtDNA) genes such as 12S rRNA and cytochrome b, which are commonly used due to their sequence diversity and the presence of multicopy genes [14]. In a previous study, the cytochrome b fragment of the mtDNA gene was used as the target to distinguish the species origin of meat samples.…”