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

Zsolnai, A. and Szántó-Egész, R. and Ferencz-Elblinger, E. and Dang Huu, A. and Jánosi, A. and Koppányné Szabó, E. and Anton, I. (2017) Loop-mediated isothermal amplification based approach as an alternative to recombinase polymerase amplification based detection of Mangalitza component in food products. Acta Alimentaria, 46 (3). pp. 384-389. ISSN 0139-3006

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Abstract

We used an alternative approach, loop-mediated isothermal amplification, to detect Mangalitza component in food products, and it has been compared to an established Recombinase Polymerase Amplification test. The correlation between the assays was significant (P<0.01). Linear determination coefficient 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 (<sc>Szántó-Egész</sc> et al., 2013) for detection of Mangalitza meat in food products, using a Mangalitza specific sequence. Other Mangalitza specific sequences suitable for the same purpose are also in use (V. <sc>Stéger</sc>, personal communication). Approaches like real-time monitoring of accumulation of the specific DNA product usually require specialised laboratory equipment. For Mangalitza detection, portable Recombinase Polymerase Amplification (RPA) approach has been developed (<sc>Szántó-Egész</sc> et al., 2016), which requires a device capable of maintaining 39 °C and a lateral flow strip with easy yes/no indication of the successful amplification. 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.

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