Short Communication: Detection of Salmonella typhimurium ATCC 14028 and Listeria monocytogenes ATCC 7644 in processed meat products using Real-Time PCR Multiplex Method

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ALFI SOPHIAN
https://orcid.org/0000-0002-5206-2110
RATNA PURWANINGSIH
EKA PUTRI JUNIARTI IGIRISA
MUHAMMAD LUTHFI AMIRULLAH
BERTHA LOLO LUKITA
RISKA ADRIANI FITRI

Abstract

Abstract. Sophian A, Purwaningsih R, Igirisa RPJ, Amirullah ML, Lukita BL, Fitri RA. 2020. Short Communication: Detection of Salmonella typhimurium ATCC 14028 and Listeria monocytogenes ATCC 7644 in processed meat products using Real-Time PCR Multiplex Method. Asian J Nad Prod Biochem 21: 17-20. The detection of Salmonella typhimurium ATCC 14028 and Listeria monocytogenes ATCC 7644 in processed meat products was carried out using Multiplex Real-Time PCR (qPCR) in the Microbiology and Molecular Biology Laboratory at the Indonesian Food and Drug Authority in Gorontalo. The purpose of this study was to provide alternative testing methods for food products circulating in the market. The sample consisted of 25 samples of processed meat products spike with Salmonella typhimurium ATCC 14028 phase 2 and Listeria monocytogenes ATCC 7644 phase 2. The method used in the study was qPCR analysis using the SYBR Green method, while DNA isolation used the direct PCR method. Data analysis was carried out based on Cycle threshold and Melting temperature based on two main criteria. Cycle threshold (Ct) analysis determines the Ct value of the sample and comparing it with the control. Melting temperature (Tm) analysis determines the temperature at which 50% of double-stranded DNA changed to a single standard and comparing it with the melting temperature of positive control. The results showed Salmonella typhimurium ATCC 14028 in the processed meat was detected at an average Ct value of 10.34, and a Tm value of 85.70. The presence of Listeria monocytogenes ATCC 7644 in the samples was recognized at an average Ct value of 14.04, and an average Tm value of 80.07. It can be concluded that the real-time multiplex PCR method can be used to detect Salmonella typhimurium ATCC 14028 and Listeria monocytogenes ATCC 7644 by using the melting curve (Tm) analysis.

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References
BPOM. 2019. Peraturan Badan Pengawas Obat dan Makanan Nomor 13 tahun 2019. Tentang Batas Cemaran Mikroba Badan Pengawas Obat dan Makanan Republik Indonesia. Jakarta. Indonesia.
Chamberlain, J. S., Gibbs, R. A., Ranier, J. E., Nguyen, P. N., dan Caskey, C. T. 1988. Deletion Screening of the Duchenne Muscular Dystrophy Locus Via Multiplex DNA Amplification. Nucleid Acid Research. 16(23): 11141-11156.
Gosiewski T, Monika B, Heczko. P.B. 2012. The application of multiplex PCR to detect seven different DNA targets in group B streptococci. Folia Microbiol (2012) 57:163–167.
Hayden, M. J., Nguyen, T. M., Waterman, A., dan Chalmers, K. J. 2008. Multiplex-ready PCR: A New Methode for Multiplexed SSR and SNP Genotyping. BMC Genomics. 9: 80.
McLauchlin, J., G. L. Narayanan, V. Mithani, and G. O. Neill. 2000. The detection of enterotoxin and toxic shock syndrome toxin genes in Staphylococcus aureus by polymerase chain reaction. J. Food Prot. 63:479–488.
Molina F, Acedo E.L. Rafael T, Isidro R, Antonia G, Jose R. 2015. Improved detection of Escherichia coli and coliform bacteria by multiplex PCR. Molina et al. BMC Biotechnology (2015) 15:48.
Oliveira ACS, Matheus C Rosa, Jéssica LB, Yasmine AM, Milena MAF, Gabrielle Virgínia FC, Andréia SS, Roberta SS, Josyane BS, Fábio FL, Talita BR, Carina M. 2018. Validating the Efficiency of a Simplex PCR and Quantitative SYBR Green qPCR for the Identification of Salmonella spp. DNA. J Food Microbiol Saf Hyg. 3:1.
Sophian A. Purwaningsih R, Lukita B L And Cahyaningsih E. 2020. Detection of Salmonella typhimurium ATCC 14028 in supplement health product liquid preparation using Real-Time PCR (qPCR). Biofarmasi J Nat Prod Biochem 18: 61-65.
Sugiyoto, Kusuma A, & Veronica W. 2015. Kandungan mikroba pada daging sapi dari beberapa pasar tradisional di Bandar Lampung. Jurnal Ilmiah Peternakan Terpadu Vol. 3(2): 27-30, Mei 2015.