Combination of Loop-Mediated Isothermal Amplification and AuNP-Oligoprobe Colourimetric Assay for Pork Authentication in Processed Meat Products

Abstract

Pork adulteration is a major concern for Muslims and Jews whose diets are restricted by religious beliefs, as well as those who are allergic to pork meat and its derivatives. Accurate pork authentication is of great importance to assist this demographic group of people in making decision on their product purchase. The aim of this study was to develop a new analytical method for pork authentication in processed meat products based on a combination of loop-mediated isothermal amplification (LAMP) and AuNP-nanoprobe colourimetric assay. The LAMP conditions were first optimised to obtain the highest yield of amplified DNA products within the shortest time. Oligoprobe-functionalised AuNPs were then hybridised with LAMP-DNA amplicons and subsequently challenged with MgSO4 at a high concentration to induce AuNP aggregation. In the presence of pork DNA, the colloidal AuNP-probe remained unchanged in its red colour, which indicates the dispersion of AuNPs. In contrast, in the absence of pork DNA, the colour was changed to colourless as a result from the aggregation of AuNPs. The LAMP-AuNP-nanoprobe assay offers a high sensitivity with a limit of detection as low as 100 pg of pork DNA. The assay is highly specific to pork content without cross-reactivity with the other meat species tested. The assay developed herein can become a simple, inexpensive, precise, and rapid analytical tool for small laboratories or the general public interested in halal food authentication.

Loop-Mediated Isothermal Amplification (LAMP): The Better Sibling of PCR?

“…Food frauds are especially prevalent in poor and developing countries with high populations, where the demand for meat and meat products is rising, which inflates their prices [63]. Subject literature contains an increasing number of reports on pork frauds and their detectability with LAMP [64][65][66][67][68].…”

The Utilization of the SaLux19 -Based LAMP Assay for the Rapid and Sensitive Identification of Minute Amounts of Biological Material

“…The sample for this study was selected according to the following criteria: Pork and its derivates are religiously forbidden for some groups, such as Jews and Muslims [8,9]. Moreover, the consumption of pig tissue-related products can trigger the development of allergies in sensitive individuals [9,10].…”

Rapid Full-Cycle Technique to Control Adulteration of Meat Products: Integration of Accelerated Sample Preparation, Recombinase Polymerase Amplification, and Test-Strip Detection

“…To control the adulteration of meat products, approaches have been proposed based on different variants of isothermal amplification, and attention to these techniques has increased in recent years [27,35]. Loop-mediated isothermal amplification (LAMP) is the most popular method for rapid DNA detection [36][37][38][39][40][41]. These tests utilize fluorescence detection or color changing visualization to enable naked-eye detection.…”