Loop-mediated isothermal amplification based approach as an alternative to recombinase polymerase amplification based detection of Mangalitza component in food products

Abstract

We used an alternative approach, loop-mediated isothermal amplifi cation, to detect Mangalitza component in food products, and it has been compared to an established Recombinase Polymerase Amplifi cation test. The correlation between the assays was signifi cant (P<0.01). Linear determination coeffi cient between the assays was 0.993 and level of diagnostic agreement was high (Kappa=0.971).Previously, a real-time PCR method based on TaqMan probe was developed for detection of Mangalitza meat in food products, using a Mangalitza specifi c sequence. Other Mangalitza specifi c sequences suitable for the same purpose are also in use (V. STÉGER, personal communication).Approaches like real-time monitoring of accumulation of the specifi c DNA product usually require specialised laboratory equipment. For Mangalitza detection, portable Recombinase Polymerase Amplifi cation (RPA) approach has been developed (SZÁNTÓ-EGÉSZ et al., 2016), which requires a device capable of maintaining 39 °C and a lateral fl ow strip with easy yes/no indication of the successful amplifi cation.We wanted to develop another fast, non-PCR based test with minimal laboratory requirement to provide a third possibility to detect Mangalitza component in food.Keywords: food, LAMP, RPA, MangalitzaNowadays, a wide range of non-PCR amplifi cations are available, such as Helicase Dependent Amplifi cation (VINCENT et al., 2004), Transcription Mediated Amplifi cation (GUATELLI et al., 1990), Self-Sustained Sequence Replication (3SR) (GUATELLI et al., 1990), Rolling Circle Amplifi cation (FIRE and XU, 1995), Standard Displacement Amplifi cation (SDA) (WALKER et al., 1992), and loop-mediated isothermal amplifi cation (LAMP) (NOTOMI et al., 2000). They usually do not require expensive instrumentation and have comparable diagnostic power to * To whom correspondence should be addressed. E-mail: attila.zsolnai@gmail.com This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (https://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted use, distribution, and reproduction in any medium for non-commercial purposes, provided the original author and source are credited, a link to the CC License is provided, and changes -if any -are indicated.

Genetic position of Hungarian Grey among European cattle and identification of breed-specific markers

“…SNPs performing well in differentiating Hungarian Greys from other breeds ( Figure 5) are the base of developing molecular trademark (Szántó-Egész et al, 2016;Zsolnai et al, 2017) for the beef produced by Hungarian Grey breeders. Similar achievements have been described in cattle such as the inclusion of seven breeds into the quest for breed-specific SNPs (Czech et al, 2018); testing of different identification methods of a small, cost-effective set of SNPs for breed assignment (Kumar et al, 2019); or finding trait-associated loci (Mastrangelo et al, 2016).…”

“…The objectives of the present paper were to compare the genetic status of Hungarian Grey to other European cattle in the hope to get insight into the history of the breed, like in the case of Maltese (Lancioni et al, 2016), and to initialise molecular trademark development, already performed in Mangalitza pig (Szántó-Egész et al, 2016;Zsolnai et al, 2017), via looking for differences between Hungarian Grey and European cattle breeds.…”