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13 (1) 2023

Real-time PCR application in confirmation test of Salmonella Typhimurium on instant noodle


Author - Affiliation:
Alfi Sophian - National Center for Drug and Food Testing Development , Indonesia
Sri Utaminingsih - National Center for Drug and Food Testing Development , Indonesia
Tirta Setya Bhakti - Center for Drug and Food Control in Semarang , Indonesia
Dewi Rahmawati - Center for Drug and Food Control in Banjarmasin , Indonesia
Corresponding author: Alfi Sophian - alfi.sophian@pom.go.id
Submitted: 24-09-2022
Accepted: 11-01-2023
Published: 05-04-2023

Abstract
Real-Time PCR is a molecular biology testing instrument used for various types of food product testing applications to monitor the quality of a product. In addition, the real-time PCR technique can also be used to test the detection of pathogenic bacteria in some foods and processed products. The purpose of this study was to detect Salmonella typhimurium using real-time PCR. This research can be a source of information and a reference for similar research. The test method used is the real-time PCR method with the green SYBR kit. Data analysis was carried out by looking at the results of interpreting Ct values and Tm values. Positive results are indicated by the formation of the Ct curve and the Tm curve. From the research conducted, it was found that the Ct value was in the range of 11.90 - 12.10 with an average of 12.02, while the Tm value was in the range of 85.10 - 85.20 with an average of 85.15. From the study results, it can be concluded that the use of real-time PCR for testing the detection of Salmonella typhimurium in instant noodle products can be applied.

Keywords
bacteria; DNA; PCR; salmonella; testing

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Sophian, A., Utaminingsih, S., Bhakti, T. S., Rahmawati, D. (2023). Real-time PCR application in confirmation test of Salmonella Typhimurium on instant noodle. Ho Chi Minh City Open University Journal of Science – Engineering and Technology, 13(1), 3-9. doi:10.46223/HCMCOUJS.tech.en.13.1.2479.2023


References

Beutin, L., Jahn, S., & Fach, P. (2009). Evaluation of the ‘GeneDisc’ real-time PCR system for detection of enterohaemorrhagic Escherichia coli (EHEC) O26, O103, O111, O145 and O157 strains according to their virulence markers and their O- and H-antigen-associated genes. Journal of Applied Microbiology, 106(4), 1122-1132.


Carlin, C. R., Lau, S. S., Cheng, R. A., Buehler, A. J., Kassaify, Z., & Wiedmann, M. (2020). Validation using diverse, difficult-to-detect salmonella strains and a dark chocolate matrix highlights the critical role of strain selection for evaluation of simplified, rapid PCR-based methods offering next-day time to results. Journal of Food Protection, 83(8), 1374-1386.


Ding, T., Suo, Y., Zhang, Z., Liu, D., Ye, X., Chen, S., & Zhao, Y. (2017). A multiplex RT-PCR assay for S. aureus, L. monocytogenes, and Salmonella spp. Detection in raw milk with pre-enrichment. Frontiers in Microbiology, 8, 1-11.


Dwight, Z., Palais, R., & Wittwer, C. T. (2011). uMELT: Prediction of high-resolution melting curves and dynamic melting profiles of PCR products in a rich web application. Bioinformatics, 27(7), 1019-1020.


Fukushima, H., Katsube, K., Tsunomori, Y., Kishi, R., Atsuta, J., & Akiba, Y. (2009). Comprehensive and rapid real-time PCR analysis of 21 foodborne outbreaks. International Journal of Microbiology, 2009, 1-13.


Giacomazzi, S., Leroi, F., & Joffraud, J.-J. (2005). Comparison of three methods of DNA extraction from cold-smoked salmon and impact of physical treatments. Journal of Applied Microbiology, 98(5), 1230-1238.


Greve-Peterson, J., & Legan, J. D. (n.d.). Evaluation of rapid screening platforms for detection of salmonella in probiotic cultures. Retrieved May 10, 2022, from https://cdnmedia.eurofins.com/eurofins-us/media/1709578/7965720-pos-866233-01b.pdf


Ibrahim, W. A., El-Ghany, W. A. A., Nasef, S. A., & Hatem, M. E. (2014). A comparative study on the use of Real Time Polymerase Chain Reaction (RT-PCR) and standard isolation techniques for the detection of Salmonellae in broiler chicks. International Journal of Veterinary Science and Medicine, 2(1), 67-71.


Jobling, M. A., & Gill, P. (2004). Encoded evidence: DNA in forensic analysis. Nature Reviews Genetics, 5(10), 739-751.


Lockley, A. K., & Bardsley, R. G. (2000). DNA-based methods for food authentication. Trends in Food Science & Technology, 11(2), 67-77.


Malorny, B., Hoorfar, J., Bunge, C., & Helmuth, R. (2003). Multicenter validation of the analytical accuracy of Salmonella PCR: Towards an international standard. Applied and Environmental Microbiology, 69(1), 290-296.


Mata, G. M. S. C., & Vanetti, M. C. D. (2012). Comparison of conventional and rapid methods for Salmonella detection in Artisanal Minas Cheese. Journal of Food Research, 1(3), 178-183.


Paniel, N., & Noguer, T. (2019). Detection of Salmonella in food matrices, from conventional methods to recent aptamer-sensing technologies. Foods, 8(9), 371-407.


Postollec, F., Falentin, H., Pavan, S., Combrisson, J., & Sohier, D. (2011). Recent advances in quantitative PCR (qPCR) applications in food microbiology. Food Microbiology, 28(5), 848-861.


Sapiun, Z., Sophian, A., Abinawanto, Muindar, Kamba, V., Damiti, S. A., & Luawo, H. (2020). Optimization of Mcfarland Turbidity standards value in determining template DNA as reference in Salmonella Typhimurium ATCC 14028 test using Real-Time PCR (QPCR). PalArch’s Journal of Archaeology of Egypt/Egyptology, 17(6), 10916-10922.


Siala, M., Barbana, A., Smaoui, S., Hachicha, S., Marouane, C., Kammoun, S., … Messadi-Akrout, F. (2017). Screening and detecting Salmonella in different food matrices in Southern Tunisia using a combined enrichment/Real-Time PCR method: Correlation with conventional culture method. Frontiers in Microbiology, 8, 1-10.


Sophian, A., Purwaningsih, R., Igirisa, R. P. J., Amirullah, M. L., Lukita, B. L., & Fitri, R. A. (2021). Short communication: Detection of salmonella typhimurium ATCC 14028 and listeria monocytogenes ATCC 7644 in processed meat products using Real-Time PCR multiplex method. Asian Journal of Natural Product Biochemistry, 19, 17-20. 


Sophian, A., Purwaningsih, R., Lukita, B. L., & Ningsih, E. C. (2020). Detection of Salmonella typhimurium ATCC 14028 in supplement health product liquid preparation using Real-Time PCR (qPCR). Biofarmasi Journal of Natural Product Biochemistry, 18(2), 65-69.


Sophian, A., Purwaningsih, R., Muindar, M., Igirisa, E. P. J., & Amirullah, M. L. (2021a). Detection of Salmonella typhimurium ATCC 14028 in powder prepared traditional medicines using Real-Time PCR. Borneo Journal of Pharmacy, 4(3), 178-183. 


Sophian, A., Purwaningsih, R., Muindar, M., Igirisa, E. P. J., & Amirullah, M. L. (2021b). Use of direct PCR technique without DNA extraction in confirmation test for Salmonella typhimurium bacteria on meatball samples. Borneo Journal of Pharmacy, 4(4), 324-332.


Sturza, R., Mitin, V., Mitina, I., Buga, A., Zgardan, D., & Behta, E. (2021). Development and Validation of SYBR Green-Based qPCR technique of detection and quantification of Salmonella enterica. Food and Nutrition Sciences, 12(11), 997-1007.


Tang, Y.-W., Procop, G. W., & Persing, D. H. (1997). Molecular diagnostics of infectious diseases. Molecular Diagnostics of Infectious Diseases, 43(11), 2021-2038.


Wilson, I. G. (1997). Inhibition and facilitation of nucleic acid amplification. Applied and Environmental Microbiology, 63(10), 3741-3751.


Zhao, L., Wang, J., Sun, X. X., Wang, J., Chen, Z., Xu, X., … Geng, Y. (2021). Development and Evaluation of the Rapid and Sensitive RPA Assays for Specific Detection of Salmonella spp. In Food Samples. Frontiers in Cellular and Infection Microbiology, 11, Article 631921.



DOI: https://doi.org/10.46223/HCMCOUJS.tech.en.13.1.2479.2023
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