Browse > Article
http://dx.doi.org/10.13103/JFHS.2021.36.4.289

Development of Raw Material Identification Method of Changnan-jeot and Gaiyang-jeot Using Multiplex PCR and Real-Time PCR  

Choi, Seong Seok (Basic Science Research Institute, Pukyong National University)
Seo, Yong Bae (Basic Science Research Institute, Pukyong National University)
Kim, Jong-Oh (Institute of Marine Biotechnology, Pukyong National University)
Yang, Ji-Young (Department of Food Science & Technology, Pukyoung National University)
Shin, Jiyoung (Institute of Food Science, Pukyong National University)
Kim, Gun-Do (Department of Microbiology, Pukyong National University)
Publication Information
Journal of Food Hygiene and Safety / v.36, no.4, 2021 , pp. 289-297 More about this Journal
Abstract
In this study, multiplex PCR and real-time PCR were performed on Theragra chalcogramma (walleye pollock), Pangasianodon hypophthalmus (iridescent shark) and their processed foods, such as changnan-jeot and gaiyang-jeot (salted iridescent shark intestine). Species-specific primers for T. chalcogramma and P. hypophthalmus were designed, and genomic DNA was directly extracted from each sample to perform single PCR and multiplex PCR. As a result of PCR, in the case of single PCR, PCR bands of T. chalcogramma (297 bp) and P. hypophthalmus (132 bp) were identified, and in the case of multiplex PCR, it was confirmed that amplification occurred without cross-reaction between T. chalcogramma and P. hypophthalmus. As a result of checking the PCR sensitivity, the concentration of genomic DNA was detected up to 0.1 ng/µL in both single PCR and multiplex PCR. The real-time PCR results showed that the average Ct value of T. chalcogramma was 20.765±0.691, and the average Ct value of P. hypophthalmus sample was 35.719±1.828 in the T. chalcogramma species-specific primers. In the P. hypophthalmus species-specific primers, the average Ct value of the T. chalcogramma sample was 35.996±1.423, and the mean Ct value of the P. hypophthalmus sample was 20.096±0.793. These results demonstrated the significant differences in the efficiency, specificity and cross-reactivity of species-specific primers in real-time PCR. Based on these findings, 7 of changnan-jeot or gaiyang-jeot products were confirmed by multiplex PCR and real-time PCR, and valid results were confirmed in all samples.
Keywords
Multiplex PCR; Real-time PCR; Changnan-jeot; Theragra chalcogramma; Pangasianodon hypophthalmus;
Citations & Related Records
Times Cited By KSCI : 6  (Citation Analysis)
연도 인용수 순위
1 Chung, I.Y., Seo, Y.B., Yang, J.Y., Kim, G.D., Development and validation of Real-time PCR to determine Branchiostegus japonicus and B. albus species based on mitochondrial DNA. J. Life Sci., 27, 1331-1339 (2017).   DOI
2 Ministry of Food and Drug Safety: Guidelines for determining the authenticity of ingredients used in food (II) Utilization of genetic analysis method, pp. 16-90. (2012).
3 Ministry of Food and Drug Safety: Genetic analysis method to determine the authenticity of raw materials used in food, pp. 103-113 (2016).
4 Aida, A.A., Che Man, Y.B., Wong, C.M.V.L., Raha, A.R., Son, R., Analysis of raw meats and fats of pig using polymerase chain reaction for halal authentication. Meat Sci., 69, 47-52 (2005).   DOI
5 Yuan, J.S., Reed, A., Chen, F., Stewart Jr, C.N., Statistical analysis of real-time PCR data. BMC Bioinformatics, 7, 85 (2006).   DOI
6 Kim, K,H, Kim, M.R., Park, Y.E., Kim, Y.S., Lee, H.Y., Park, Y.C., Kim, S.Y., Choi, J.D., Jang, Y.M., Detection and differentiation of intentional and unintentional mixture in raw meats using real-time PCR. J. Food Hyg. Saf., 29, 340-346 (2014).   DOI
7 Birky-Jr, C.W., Fuerst, P., Maruyamat, T., Organelle gene diversity under migration, mutation, and drift: equilibrium expectations, approach to equilibrium, effects of heteroplasmic cells, and comparison to nuclear genes. Genetics, 121, 613-627 (1989).   DOI
8 Ha, J.C., Jung, W.T., Nam, Y.S., Moon, T.W., PCR identification of ruminant tissue in raw and heat-treated meat meals. J. Food Prot., 69, 2241-2247 (2006).   DOI
9 Park, Y.C., Jin, S.O., Lim, J.Y., Kim, K.H., Lee, J.H., Cho, T.Y., Lee, H.J., Han, S.B., Lee, S.J., Lee, K.H., Yoon, H.S., Application for identification of food raw materials by PCR using universal primer. J. Food Hyg. Saf., 27, 317-324 (2012).   DOI
10 Lin, X., Kim, K.S., Empirical analysis on the factors affecting the consumption of aquatic products in China using panel data. J. Fish. Bus. Adm., 44, 19-33 (2013).   DOI
11 Anderson, S., Bankier, A.T., Barrell, B.G., de Bruijn, M.H.L., Coulson, A.R., Drouin, J., Eperon, I.C., Nierlich, D.P., Roe, B.A., Sanger, F., Schreier, P.H., Smith, A.J.H., Staden, R., Young, I.G., Sequence and organization of the human mitochondrial genome. Nature, 290, 457-465 (1981).   DOI
12 Irwin, D.M., Kocher, T.D., Wilson, A.C., Evolution of the cytochrome b gene of mammals. J. Mol. Evol., 32, 128-144. (1991).   DOI
13 Park, J.K., Shin, K.H., Shin, S.C., Chung, K.Y., Chung, E.R., Identification of meat species using species-specific PCR-RFLP fingerprint of mitochondrial 12S rRNA gene. Food Sci. Anim. Resour., 27, 209-215 (2007).   DOI
14 Sezaki, K., Itoi, S., Watabe, S., A simple method to distinguish two commercially valuable eel species in Japan Anguilla japonica and A. anguilla using polymerase chain reaction strategy with a species-specific primer. Fish. Sci., 71, 414-421 (2005).   DOI
15 Tanabe, S., Miyauchi, E., Muneshige, A., Mio, K., Sato, C., Sato, M., PCR method of detecting pork in foods for verifying allergen labeling and for identifying hidden pork ingredients in processed foods. Biosci. Biotechnol. Biochem., 71, 1663-1667 (2007).   DOI
16 Lee, W.D., Chang, D.S., Kang, S.M., Yoon, J.H., Lee, M.S., Development of new manufacturing process for Changranjeotgal. Korean J. Fish. Aquat. Sci., 34, 109-113 (2001).
17 Espineira, M., Vieites, J.M., Santaclara, F.J., Species authentication of octopus, cuttlefish, bobtail and bottle squids (families Octopodidae, Sepiidae and Sepiolidae) by FINS methodology in seafoods. Food Chem., 121, 527-532 (2010).   DOI
18 Santaclara F.J., Espineira, M., Vieites, J.M., Genetic Identification of Squids (Families Ommastrephidae and Loliginidae) by PCR-RFLP and FINS Methodologies. J. Agric. Food Chem., 55, 9913-9920 (2007).   DOI
19 Chapela, M.J., Sotelo, C.G., Calo-Mata, P., Perez-Martin, R.I., Rehbein, H., Hold, G.L., Quinteiro, J., Rey-Mendez, M., Rosa, C., Santos, A.T., Identification of cephalopod species (Ommastrephidae and Loliginidae) in seafood products by forensically informative nucleotide sequencing (FINS). J. Food Sci., 67, 1672-1676 (2006).   DOI
20 Kang, S.K., Park, J.H., Kim, S.A., Size-class estimation of the number of walleye pollock Theragra chalcogramma caught in the Southwestern East Sea during the 1970s-1990s. Korean J. Fish. Aquat. Sci., 46, 445-453 (2013).   DOI
21 Seo, Y.B., Kang, S.C., Choi, S.S., Lee, J.K., Jeong, T.H., Lim, H.K., Kim, G.D., Phylogenetic study of genus Haliotis in Korea by cytochrome c oxidase subunit 1 and RAPD analysis. J. Life Sci., 26, 406-413. (2016).   DOI
22 Kim, H.S., Seo, Y.B., Choi, S.S., Kim, J.H., Shin, J.Y., Yang J.Y., Kim, G.D., Development and validation of multiplex polymerase chain reaction to determine squid species based on 16s rRNA gene. J. Food Hyg. Saf., 30, 43-50 (2015).   DOI
23 Martin, I., Garcia, T., Fajardo, V., Lopez-Calleja, I., Rojas, M., Pavon, M.A., Hernandez, P.E., Gonzalez, I., Martin, R., Detection of chicken, turkey, duck, and goose tissues in feed-stuffs using species-specific polymerase chain reaction. J. Anim. Sci., 85, 452-458 (2007).   DOI