Polymerase chain reaction (PCR) has become the main method
for detection and identification of genetically modified organisms (GMOs). Multiplex
PCR is a variation of this technique allowing simultaneous amplification of
several targets in the same reactions. In this study, it was developed, optimized
and performed in-house validation of the multiplex PCR method for detection
and identification of genetically modified rape lines (GT73, Ms8, Rf3 and T45) in
two types of assays: gene- and event-specific. The optimized reactions exhibited
high specificity, sensitivity, selectivity and accuracy. The limit of detection (LOD)
for the gene-specific reaction was 0.01% for each of the tested lines of rape. For
the event-specific reaction, LOD was at the same level for most of the GM rape
lines except for T45 (LOD = 0.025%). The optimized and validated assays were
employed from 2012 to 2015 in our laboratory to analyse 428 samples of animal
feedstuffs containing oilseed rape. GM rape was not detected in the analysed
feedstuffs until 2013 (only some traces of GM soybean were found). Since 2014,
the GM rape presence has been confirmed in event-specific reactions in more
than one third of analysed samples.
REFERENCES(30)
1.
Datukishvili N., Kutateladze T., Gabriadze I., Bitskinashvili K., Vishnepolsky B., 2015. New multiplex PCR methods for rapid screening of genetically modified organisms in foods. Front. Microbiol. 6, 757, https://doi.org/10.3389/fmicb.....
Demeke T., Giroux R.W., Reitmeier S., Simon S.L., 2002. Development of a polymerase chain reaction assay for detection of three canola transgenes. J. Am. Oil Chem. Soc. 79, 1015–1019, https://doi.org/10.1007/s11746....
Demeke T., Ratnayaka I., 2008. Multiplex qualitative PCR assay for identification of genetically modified canola events and real-time event-specific PCR assay for quantification of the GT73 canola event. Food Control 19, 893–897, https://doi.org/10.1016/j.food....
Dobnik D., Štebih D., Blejec A., Morisset D., Žel J., 2016. Multiplex quantification of four DNA targets in one reaction with Bio-Rad droplet digital PCR system for GMO detection. Sci. Rep. 6, 35451, https://doi.org/10.1038/srep35....
Dörries H.-H., Remus I., Grönewald A., Grönewald C., Berghof-Jäger K., 2010. Development of a qualitative, multiplex realtime PCR kit for screening of genetically modified organisms (GMOs). Anal. Bioanal. Chem. 396, 2043–2054, https://doi.org/10.1007/s00216....
EN ISO 21571:2005. Foodstuffs – Methods of analysis for the detection of genetically modified organisms and derived products – Nucleic acid extraction. https://www.iso.org/standard/3....
Fernandes T.J.R., Amaral J.S., Oliveira M.B.P.P., Mafra I., 2014. A survey on genetically modified maize in foods commercialised in Portugal. Food Control 35, 338–344, https://doi.org/10.1016/j.food....
Forte V.T., Di Pinto A., Martino C., Tantillo G.M., Grasso G., Schena F.P., 2005. A general multiplex-PCR assay for the general detection of genetically modified soya and maize. Food Control 16, 535–539, https://doi.org/10.1016/j.food....
Germini A., Zanetti A., Salati C., Rossi S., Forré C., Schmid S., Marchelli R., 2004. Development of a seven-target multiplex PCR for the simultaneous detection of transgenic soybean and maize in feeds and foods. J. Agric. Food Chem. 52, 3275–3280, https://doi.org/10.1021/jf0350....
Hellebrand M., Nagy M., Mörsel J.-T., 1998. Determination of DNA traces in rapeseed oil. Z. Lebensm. Unters. Forsch. 206, 237–242, https://doi.org/10.1007/s00217....
Holst-Jensen A., Bertheau Y., de Loose M. et al., 2012. Detecting unauthorized genetically modified organisms (GMOs) and derived materials. Biotechnol. Adv. 30, 1318–1335, https://doi.org/10.1016/j.biot....
Holst-Jensen A., Ronning S.B., Lovseth A., 2003. PCR technology for screening and quantification of genetically modified organisms (GMOs). Anal. Bioanal. Chem. 375, 985–993, https://doi.org/10.1007/s00216....
Iwobi A., Gerdes L., Busch U., Pecoraro S., 2016. Droplet digital PCR for routine analysis of genetically modified foods (GMO) – A comparison with real-time quantitative PCR. Food Control 69, 205–213, https://doi.org/10.1016/j.food....
James D., Schmidt A.-m., Wall E., Green M., Masri S., 2003. Reliable detection and identification of genetically modified maize, soybean and canola by multiplex PCR analysis. J. Agric. Food Chem. 51, 5829–5834, https://doi.org/10.1021/jf0341....
Kim J.-H., Kim T.-W., Lee W.-Y., Park S.-H., Kim H.-Y., 2007. Multiplex PCR detection of the GT73, MS8xRF3, and T45 varieties of GM canola. Food Sci. Biotechnol. 16, 104–109.
Kim J.-H., Zhang D., Kim H.-Y., 2014. Detection of sixteen genetically modified maize events in processed foods using four eventspecific pentaplex PCR systems. Food Control 35, 345–353, https://doi.org/10.1016/j.food....
Marmiroli N., Maestri E., Gullì M., Malcevschi A., Peano C., Bordoni R., De Bellis G., 2008. Methods for detection of GMOs in food and feed. Anal. Bioanal. Chem. 392, 369–384, https://doi.org/10.1007/s00216....
Miraglia M., Berdal K.G., Brera C. et al., 2004. Detection and traceability of genetically modified organisms in the food production chain. Food Chem. Toxicol. 42, 1157–1180, https://doi.org/10.1016/j.fct.....
Özgen Arun Ö., Yılmaz F., Muratoğlu K., 2013. PCR detection of genetically modified maize and soy in mildly and highly processed foods. Food Control 32, 525–531, https://doi.org/10.1016/j.food....
Santa-Maria M.C., Lajo-Morgan G., Guardia L., 2014. Adventitious presence of transgenic events in the maize supply chain in Peru: a case study. Food Control 41, 96–101, https://doi.org/10.1016/j.food....
Shin K.-S., Suh S.-C., Lim M.-H., Woo H.-J., Lee J.H., Kim H.-Y., Cho H.-S., 2013. Event-specific detection system of stacked genetically modified maize by using the multiplex-PCR technique. Food Sci. Biotechnol. 22, 1763–1772, https://doi.org/10.1007/s10068....
Turkec A., Lucas S.J., Karlık E., 2016. Monitoring the prevalence of genetically modified maize in commercial animal feeds and food products in Turkey. J. Sci. Food Agric. 96, 3173–3179, https://doi.org/10.1002/jsfa.7....
Wu G., Wu Y., Xiao L., Lu C., 2008. Event-specific qualitative and quantitative polymerase chain reaction methods for detection of genetically modified rapeseed Ms8Rf3 based on the right border junctions. J. AOAC Int. 91, 143–151.
Yang L., Pan A., Zhang H., Guo J., Yin C., Zhang D., 2006. Event-specific qualitative and quantitative polymerase chain reaction analysis for genetically modified canola T45. J. Agric. Food Chem. 54, 9735–9740, https://doi.org/10.1021/jf0619....
Yang R., Xu W., Luo Y., Guo F., Lu Y., Huang K., 2007. Event-specific qualitative and quantitative PCR detection of roundup ready event GT73 based on the 3’-integration junction. Plant Cell Rep. 26, 1821–1831, https://doi.org/10.1007/s00299....
Zdjelar G., Nikolić Z., Vasiljević I., Bajić B., Jovičić D., Ignjatov M., Milošević D., 2013. Detection of genetically modified soya, maize, and rice in vegetarian and healthy food products in Serbia. Czech J. Food Sci. 31, 43–48.
Zeitler R., Pietsch K., Waiblinger H.-U., 2002. Validation of real-time PCR methods for the quantification of transgenic contaminations in rape seed. Eur. Food Res. Technol. 214, 346–351, https://doi.org/10.1007/s00217....
Development of ultrafast PCR assays for the event-specific detection of eleven approved genetically modified canola events in South Korea Ji-Eun Park, Do-Geun Lee, Hong-Rae Kim, Mi-Ju Kim, Hae-Yeong Kim, Hyun-Joong Kim Food Chemistry
We process personal data collected when visiting the website. The function of obtaining information about users and their behavior is carried out by voluntarily entered information in forms and saving cookies in end devices. Data, including cookies, are used to provide services, improve the user experience and to analyze the traffic in accordance with the Privacy policy. Data are also collected and processed by Google Analytics tool (more).
You can change cookies settings in your browser. Restricted use of cookies in the browser configuration may affect some functionalities of the website.