Comparison of real time PCR and conventional PCR by identifying genomic DNA of bovine and porcine

Muhammad Abdurrahman Munir,Ahlam Inayatullah

doi:10.14203/inajac.v23i2.491

Abstract

Bovine and porcine are poultry meat that consumed worldwide particularly in Southeast Asia.Both of them are prone to food counterfeit owing to several factors such as price, appetite and Halal status. Sensitive and selective analytical methods are required to control meat products that distributed to markets. This paper studied the sensitivity between real – time and conventional PCR or known as qPCR and cPCR, respectively. Bovine and porcine were samples used to verify the sensitivity of them. Nevertheless, those instruments did not show a specific difference during DNA analysis of bovine and porcine. In conventional PCR, two pairs of DNA primers targeted cytochrome b (Cyt b) was analyzed, resulting of 120 and 131 amplicons, respectively. While qPCR applied to analyze porcine and bovine DNA. The detection limit of qPCR after porcine and bovine analysis were at 0.004 and 0.007 µg/µL, respectively. Results demonstrated the qPCR was reliable for verifying porcine and bovine DNA compared to conventional PCR. Furthermore, the study concluded that the developed assay can be easily employed for the identification of porcine and bovine tissue in food products in low resource areas.

Highlights

Nowadays, analysis of food products is imperative to identify the quantity and quality of food, preventing food adulteration and promoting food safety
Meat species analysis becomes a persistent issue that must be handled for several reasons such as (a) the quantity of meat that is different compared on the product label, (b) substitute high-quality meats partially, even for some cases convert entirely with low-quality put counterfeit label deliberately before distributed to markets, (c) the concentration of meat inside non – meat products and (d) to follow the regulation of certain country related to Halal food, where Islamic law stringently prohibits the consumption of specific meat products (Dolch et al 2020; Zia et al 2020; Kang et al 2021)
2.1.1 Sample Preparation Two different genomic DNAs animal species samples from porcine and bovine sources were obtained from Eurofins, respectively (Table 1)

Summary

Introduction

Analysis of food products is imperative to identify the quantity and quality of food, preventing food adulteration and promoting food safety. Meat species analysis becomes a persistent issue that must be handled for several reasons such as (a) the quantity of meat that is different compared on the product label, (b) substitute high-quality meats partially, even for some cases convert entirely with low-quality put counterfeit label deliberately before distributed to markets, (c) the concentration of meat inside non – meat products and (d) to follow the regulation of certain country related to Halal food, where Islamic law stringently prohibits the consumption of specific meat products (e.g., porcine products) (Dolch et al 2020; Zia et al 2020; Kang et al 2021). PCR technique is strongly selective and sensitive by multiplying nucleotide sequence-specific nucleotides exponentially in vitro (Toohey – Kurth et al 2020)

Objectives

Methods

Results

Conclusion