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Detection of Gluten using Molecular Techniques.

Niketha, N. (2014) Detection of Gluten using Molecular Techniques. [Student Project Report] (Submitted)

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Abstract

In recent years there has been growing demand for gluten free (GF) products due to the increasing prevalence of allergic reactions and intolerance, most notably celiac disease. Celiac disease (CD) is a chronic systemic autoimmune disorder caused by a permanent intolerance to gluten proteins in genetically susceptible individuals. Adherence to lifelong GF diet remains the cornerstone treatment for celiac patients. Foods that naturally do not contain wheat, barley, rye or their crossbred varieties as well as foods that contain these ingredients but have been specially processed to remove gluten are known as GF foods. Apart from diagnosed celiacs, GF diets have gained considerable popularity amongst consumers as they are perceived to be healthier than their gluten-containing counterparts. Gluten detection is of high interest for food safety of celiac patients, since the only treatment known until now is a lifelong avoidance of this protein in the diet. Thus, the availability of immunochemical and molecular methods to determine gluten markers in foods is of utmost importance to ensure the well- being of gluten-sensitive individuals. The present study deals with the development of a SYBR green based real-time PCR method allowing the detection of traces of wheat in food for celiacs. The method is based on the amplification of a sequence of the gene coding for the low molecular weight glutenin found in wheat. It allows the detection of three commonly used wheat varieties, Triticum aestivum, Triticum dicoccum and Triticum durum and does not show any cross-reactivity with the phylogenetically similar rye and barley samples. The limit of detection, determined by analysing serially diluted wheat DNA extracts, was 100pg wheat DNA. In rice- spiked model samples, the limit of detection was 0.1% wheat. The suitability of the method was evaluated by testing gluten free products. Wheat DNA was found in two out of six gluten free products. The developed Real time PCR method can be used as a supplementary tool in analysis of gluten-free and naturally gluten-free foods.

Item Type: Student Project Report
Uncontrolled Keywords: Gluten analysis, Wheat, Gluten, Polymerase chain reaction (PCR), Real Time-PCR
Subjects: 600 Technology > 08 Food technology > 01 Analysis
600 Technology > 08 Food technology > 21 Cereals > 04 Wheat
Divisions: Food Safety Analytical Quality Control Lab
Depositing User: Food Sci. & Technol. Information Services
Date Deposited: 20 Nov 2015 10:21
Last Modified: 20 Nov 2015 10:21
URI: http://ir.cftri.com/id/eprint/11993

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