Rapid porcine detection in gelatin-based highly processed products using loop mediated isothermal amplification
Document Type
Article
Publication Date
12-1-2021
Abstract
Low DNA concentration recovered from highly processed products such as gelatin and gelatin-based products renders difficulty in detecting porcine contamination using conventional PCR techniques. We documented here a porcine-specific loop-mediated isothermal amplification (LAMP) to identify porcine traces in gelatin products. The porcine-specific primers were designed according to mitochondrial DNA of Cytochrome b gene sequence. Here we used two different reaction mixtures for LAMP assay (GENIE and MYRM) against the same DNA samples extracted from gelatin products and porcine-specific primers to detect the presence of porcine DNA. The porcine-specific primers were shown to be specific only to Sus scrofa against 14 DNA of other meat species. The analytical sensitivity of the LAMP assay for porcine DNA detection is 1 pg/mu L using both GENIE (within 30 m) and MYRM (within 60 m) reaction mixtures. Analysis against 32 samples of gelatin products showed that five samples were found to contain porcine DNA; two samples out of six gelatin powder samples and three gelatin capsule samples out of nine. Out of these five positive samples, three were not labeled containing porcine gelatin. Overall, LAMP assay in this study showed an excellent specificity, sensitivity and rapidity in detection of porcine DNA in gelatin products.
Keywords
Gelatin authentication, Loop isothermal amplification, LAMP, Porcine DNA, Fluorescent dye, Real-time LAMP, Calcein
Divisions
nanocat
Funders
Universiti Putra Malaysia [GP-IPS/2018/9625800],Universiti Putra Malaysia [GP-B/2018/9658200],Ministry of Education, Malaysia [6300911]
Publication Title
Journal of Food Science and Technology
Volume
58
Issue
12
Publisher
Springer Verlag (Germany)
Publisher Location
7TH FLOOR, VIJAYA BUILDING, 17, BARAKHAMBA ROAD, NEW DELHI, 110 001, INDIA