Primer Pairs Specificity Test for Frog Meat Identification Using PCR Technique

Norman Yoshi Haryono; Rizqi Layli Khusufi; Delia Wahyu Pangesti; Evi Susanti; Rina Rifqie Mariana; Hartati Eko Wardani; Norazlinaliza Salim

doi:10.22146/ijc.83626

Primer Pairs Specificity Test for Frog Meat Identification Using PCR Technique

https://doi.org/10.22146/ijc.83626

Norman Yoshi Haryono^(1*), Rizqi Layli Khusufi⁽²⁾, Delia Wahyu Pangesti⁽³⁾, Evi Susanti⁽⁴⁾, Rina Rifqie Mariana⁽⁵⁾, Hartati Eko Wardani⁽⁶⁾, Norazlinaliza Salim⁽⁷⁾

(1) Biotechnology Study Program, Department of Applied Science, Faculty of Mathematics and Natural Sciences, Universitas Negeri Malang, Jl. Semarang No. 5, Malang 65145, Indonesia; Health and Food Centre, Institute of Research and Community Services, Universitas Negeri Malang, Jl. Semarang No. 5, Malang 65145, Indonesia
(2) Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Negeri Malang, Jl. Semarang No. 5, Malang 65145, Indonesia
(3) Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Negeri Malang, Jl. Semarang No. 5, Malang 65145, Indonesia
(4) Biotechnology Study Program, Department of Applied Science, Faculty of Mathematics and Natural Sciences, Universitas Negeri Malang, Jl. Semarang No. 5, Malang 65145, Indonesia; Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Negeri Malang, Jl. Semarang No. 5, Malang 65145, Indonesia
(5) Department of Culinary Arts, Faculty of Engineering, Universitas Negeri Malang, Jl. Semarang No. 5, Malang 65145, Indonesia
(6) Department of Public Health, Faculty of Sport Sciences, Universitas Negeri Malang, Jl. Semarang No. 5, Malang 65145, Indonesia
(7) Laboratory of Halal Science Research, Halal Products Research Institute, Universiti Putra Malaysia, 43300 UPM Serdang, Selangor, Malaysia; Centre of Foundation Studies for Agricultural Science, Universiti Putra Malaysia, 43300 UPM Serdang, Selangor, Malaysia
(*) Corresponding Author

Abstract

Halal food assurance is becoming more important with the growth of the halal industry globally. Adulteration of halal meat products using non-halal sources such as pork, dog, boar, and even frog meat has become a major problem for moslems. The purpose of this study is to initiate the method for frog meat identification using polymerase chain reaction (PCR) technique. In this study, three primer pairs (Fk1-Rk1, Fk2-Rk2, Fk3-Rk3) were analyzed for their specificity toward frog meat against other common halal meat sources such as beef, chicken, shrimp, squid, and mackerel. The visualization of DNA amplification showed that primer pair Fk1-Rk1 produced primer-dimer, thus cannot be used for this circumstance. Primer pair Fk2-Rk2 showed a better result where DNA amplicon was produced at ~100 bp for frog meat and no amplicons for other meat. Primer pair Fk3-Rk3 showed a different pattern of DNA amplification for all the meat tested, where the amplicon of frog meat was shown at ~100 bp, while the other meat showed multiple amplicons or none. In conclusion, primer pairs Fk2-Rk2 and Fk3-Rk3 showed their potential as primer pairs for frog meat identification using PCR for implementing halal food assurance, although sensitivity analysis needs to be investigated.

Keywords

halal food assurance; adulteration; frog meat; polymerase chain reaction

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DOI: https://doi.org/10.22146/ijc.83626