Detection of genetically modified food products in indian market using PCR based GMO detection kit: a pilot study

The Food Safety and Standards Authority of India (FSSAI) have not approved any genetically modified (GM) food products to be manufactured, distributed, sold/or imported in the country. Many countries across the globe are legally approved to cultivate GM crops like soybean, maize, canola, cotton seeds, etc. Many people living in urban India nowadays prefer to purchase imported food products. As a result, an increasing number of food items (without GM labels) are being imported in India. Nevertheless, these products are also easily available for buyers online. Thus, it is important to understand whether these imported food items available in the Indian market are GMO-free. The objective of this study is to check the availability of GM food products in raw and processed forms in the Indian local market through the use of conventional Polymerase Chain Reaction (PCR). The study is designed to screen for the presence of regulatory genes (35S promoter and NOS terminator) which are the most common sequences found in transgenic food products. Using the cetyl trimethyl ammonium bromide (CTAB) method, DNA was extracted from 12 food samples commercially available in the Indian market (locally and online) followed by PCR to detect the presence of GM DNA using HIMEDIA’S MBPCR055 GMO detection kit. Overall, 16.66% of the total samples were tested positive for GM DNA. Of the imported food items, 33.33% were tested positive. Products that were manufactured in the US and Netherlands were tested positive for GMOs. Their main ingredients were also soy and corn. Samples manufactured in India were GMO negative.

PERKEMBANGAN PEMANFAATAN, REGULASI, DAN METODE DETEKSI PRODUK REKAYASA GENETIKA PERTANIAN DI INDONESIA / Development of Utilization, Regulation, and Detection Methods of Agricultural Genetically Modified Products in Indonesia

<p>Genetically modified crops (GM crops) have developed very fast globally, although to date controversies over the GM crop uses are still occurring. GM crops have been planted on over 191.7 million hectare area and cultivated in 26 countries in five continents. Biosafety of GM crops both globally and domestically are guaranteed through regulations made at the level of law, government regulations, related ministrial regulation including the guidelines. In general, those regulations have been implemented, thus the biosafety of GM crop utilization is guaranteed in Indonesia. Unfortunately, although Indonesia gave a certification for released permit for drought tolerant sugarcane, it only grown in a limited areas belongs to state-owned agricultural company (PTPN XI). The country has certified 27, 7, and 16 GM events for food, feed, and seeds for environment safety, respectively. The implementation of these regulations needs a monitoring system that is equipped with facilities of GMO detection laboratory with adequate capacity. Indonesia has several such laboratories. The methods of GMO detections have developed from very basic techniques, i.e. qualitative screening to the determination of specific events that define the type of trait of GMO, even quantitative detection, both single and multiplex. Each method has its own advantages. The capacities of GMO detection laboratory in Indonesia still need to be upgraded to master the fast-developing technology. The purpose of this review is to provide information on the development of global GM crops utilization including in Indonesia and the development of regulations and detection methods with their prospects and challenges.</p><p>Keywords: Genetics, modification, regulation, detection methods</p><p> </p><p><strong>Abstrak</strong></p><p>Pemanfaatan tanaman produk rekayasa genetik (PRG) telah berkembang cepat dan mendunia walaupun sampai saat ini masih terjadi kontroversi. Luas penanamannya telah mencapai 191,7 juta ha dan ditanam oleh 26 negara di lima benua. Keamanan hayati PRG secara global maupun domestik telah dijamin oleh peraturan pada tingkat undang-undang, peraturan pemerintah, peraturan kementerian terkait, dan pedoman pelaksanaannya. Secara umum peraturan peraturan tersebut telah dijalankan sehingga keamanan hayati dari pemanfaatan PRG terjamin di Indonesia. Sayangnya di Indonesia PRG yang sudah diberi izin edar hanya ditanam secara terbatas seperti tebu toleran kekeringan di beberapa kebun milik PTPN. Indonesia juga telah memberikan sertifikat aman hayati pada beberapa varietas PRG diantaranya 27 PRG pangan, tujuh PRG pakan, dan 16 PRG benih (lingkungan). Implementasi peraturan yang telah ada memerlukan sistem pengawasan yang dilengkapi dengan fasilitas laboratorium deteksi PRG dengan kapasitas yang memadai. Indonesia telah mempunyai beberapa laboratorium tersebut. Metode deteksi PRG telah berkembang dari teknik yang sangat mendasar yaitu deteksi untuk skrining kualitatif PRG sampai teknik penentuan spesifik event yang menetapkan jenis/sifat PRG, bahkan teknik deteksi secara kuantitatif yang bersifat tunggal maupun multiplex. Metode-metode deteksi tersebut memiliki keunggulan masing-masing. Laboratorium penguji PRG di Indonesia masih perlu ditingkatkan kemampuannya dengan penguasaan teknologi yang berkembang dengan pesat. Makalah ini memberikan informasi perkembangan pemanfaatan PRG global termasuk di Indonesia dan perkembangan regulasi dan metode deteksi serta prospek dan tantangan.</p><p>Kata kunci: Genetika, rekayasa, regulasi, metode deteksi</p>

Highly Sensitive GMO Detection Using Real-Time PCR with a Large Amount of DNA Template: Single-Laboratory Validation

Current genetically modified organism (GMO) detection methods allow for sensitive detection. However, a further increase in sensitivity will enable more efficient testing for large grain samples and reliable testing for processed foods. In this study, we investigated real-time PCR-based GMO detection methods using a large amount of DNA template. We selected target sequences that are commonly introduced into many kinds of GM crops, i.e., 35S promoter and nopaline synthase (NOS) terminator. This makes the newly developed method applicable to a wide range of GMOs, including some unauthorized ones. The estimated LOD of the new method was 0.005% of GM maize events; to the best of our knowledge, this method is the most sensitive among the GM maize detection methods for which the LOD was evaluated in terms of GMO content. A 10-fold increase in the DNA amount as compared with the amount used under common testing conditions gave an approximately 10-fold reduction in the LOD without PCR inhibition. Our method is applicable to various analytical samples, including processed foods. The use of other primers and fluorescence probes would permit highly sensitive detection of various recombinant DNA sequences besides the 35S promoter and NOS terminator.