Quantification of parvalbumin in commercially important Mediterranean seafood species using real time PCR
D. Houhoula, P. Dimitriou, G. Mengjezi, V. Kyrana, V. Lougovoishttps://doi.org/10.17221/390/2014-CJFSCitation:Houhoula D., Dimitriou P., Mengjezi G., Kyrana V., Lougovois V. (2015): Quantification of parvalbumin in commercially important Mediterranean seafood species using real time PCR. Czech J. Food Sci., 33: 143-147.
The parvalbumin allergen gene was quantified in various types of seafood using RT PCR. Freshly harvested specimens from 25 species of finfish, molluscs, and crustacean shellfish, commonly consumed in the Mediterranean region, were included in the investigation. DNA was extracted using the commercial NucleoSpin Food kit. The amplification of the parvalbumin gene was performed by RT PCR. Sixteen out of the 25 species examined yielded positive amplification. Positive samples, including several species of fish (Atlantic mackerel, horse mackerel, sheepshead, red mullet, sandsmelt, pandora, saddled sea bream, gilthead sea bream, red sea bream, European sea bass, blue whiting, anchovy, sardine) and cephalopods (cuttlefish, musky octopus), exhibited largely variable thresholds differing by as much as 12 cycles. Even though equal amounts of DNA were used in PCR amplification the copy number of gene-encoded parvalbumin varied between the fish species. The assay proved to be a potential tool for the detection and label management of fish allergens in food. The studies have shown that allergic reactions to food are highly individual. For some hypersensitive patients, even trace amounts can cause life-threatening allergic reactions. The results of the present study indicate that several seafood species, commonly consumed in the Mediterranean region, may pose a threat for hypersensitive individuals.Keywords:protein quantification; Real Time PCR; seafood allergyReferences:
Bugajska-Schretter Agnes, Elfman Lena, Fuchs Thomas, Kapiotis Sonja, Rumpold Helmut, Valenta Rudolf, Spitzauer Susanne (1998): Parvalbumin, a cross-reactive fish allergen, contains IgE-binding epitopes sensitive to periodate treatment and Ca2+ depletion. Journal of Allergy and Clinical Immunology, 101, 67-74 https://doi.org/10.1016/S0091-6749(98)70195-2Chen Lingyun, Hefle Sue L., Taylor Steve L., Swoboda Ines, Goodman Richard E. (2006): Detecting Fish Parvalbumin with Commercial Mouse Monoclonal Anti-frog Parvalbumin IgG. Journal of Agricultural and Food Chemistry, 54, 5577-5582 https://doi.org/10.1021/jf060291gChoi K.Y., Hong K.W. (2007): Genomic DNA sequence of mackerel parvalbumin and a PCR test for rapid detection of allergenic mackerel ingredients in food. Food Science and Biotechnology, 16: 67–70.de Martino Maurizio, Novembre Elio, Galli Luisa, de Marco Antonella, Botarelli Patrizia, Marano E., Vierucci Alberto (1990): Allergy to different fish species in cod-allergic children: In vivo and in vitro studies. Journal of Allergy and Clinical Immunology, 86, 909-914 https://doi.org/10.1016/S0091-6749(05)80154-XELSAYED S., BENNICH H. (1975): The Primary Structure of Allergen M from Cod. Scandinavian Journal of Immunology, 4, 203-208 https://doi.org/10.1111/j.1365-3083.1975.tb02618.xFæste Christiane K., Plassen Christin (2008): Quantitative sandwich ELISA for the determination of fish in foods. Journal of Immunological Methods, 329, 45-55 https://doi.org/10.1016/j.jim.2007.09.007Gajewski K.G., Hsieh Y.H. (2009): Monoclonal antibody specific to a major fish allergen: parvalbumin. Journal of Food Protection, 72: 818–825.Griesmeier U., VÃ¡zquez-CortÃ©s S., Bublin M., Radauer C., Ma Y., Briza P., FernÃ¡ndez-Rivas M., Breiteneder H. (2010): Expression levels of parvalbumins determine allergenicity of fish species. Allergy, 65, 191-198 https://doi.org/10.1111/j.1398-9995.2009.02162.xHamada Y., Tanaka H., Ishizaki S., Ishida M., Nagashima Y., Shiomi K. (2003): Purification, reactivity with IgE and cDNA cloning of parvalbumin as the major allergen of mackerels. Food and Chemical Toxicology, 41, 1149-1156 https://doi.org/10.1016/S0278-6915(03)00074-7Hildebrandt Sabine, Garber Eric A.E. (2010): Effects of processing on detection and quantification of the parvalbumin gene in Atlantic salmon (Salmo salar). Food Chemistry, 119, 75-80 https://doi.org/10.1016/j.foodchem.2009.05.074Hilger C., Thill L., Grigioni F., Lehners C., Falagiani P., Ferrara A., Romano C., Stevens W., Hentges F. (2004): IgE antibodies of fish allergic patients cross-react with frog parvalbumin. Allergy, 59, 653-660 https://doi.org/10.1111/j.1398-9995.2004.00436.xKobayashi A., Tanaka H., Hamada Y., Ishizaki S., Nagashima Y., Shiomi K. (2006): Comparison of allergenicity and allergens between fish white and dark muscles. Allergy, 61, 357-363 https://doi.org/10.1111/j.1398-9995.2006.00966.xLehrer Samuel B., Horner W. Elliott, Reese Gerald, Taylor Steven (): Why are some proteins allergenic? Implications for biotechnology. Critical Reviews in Food Science and Nutrition, 36, 553-564 https://doi.org/10.1080/10408399609527739Li Zhenxing, Zhang Yiqun, Pawar Ravindra, Wang Guoying, Lin Hong (2011): Development of an Optimized Protein Chip for the Detection of Fish Parvalbumin Allergen. Current Analytical Chemistry, 7, 349-356 https://doi.org/10.2174/157341111797183100Sharp M.F., Lopata A.L. (2014): Fish allergy: in review. Clinical Reviews in Allergy & Immunology, 46: 258–271.Shibahara Yusuke, Uesaka Yoshihiko, Wang Jun, Yamada Shoichi, Shiomi Kazuo (2013): A sensitive enzyme-linked immunosorbent assay for the determination of fish protein in processed foods. Food Chemistry, 136, 675-681 https://doi.org/10.1016/j.foodchem.2012.08.066Shiomi K., Hamada Y., Sekiguchi K., Shimakura K., Nagashima Y. (1999): Two classes of allergens parvalbumins and higher molecular weight substances, in Japanese eel and bigeye tuna. Fish Science, 65: 943–948.Sun M., Liang C., Gao H., Lin C., Deng M. (2009): Detection of parvalbumin, a common fish allergen gene in food, by real-time polymerase chain reaction. Journal of AOAC International, 92: 234–240.