پديد آورنده :
ستايش نسب، مائده
عنوان :
طراحي آپتاسنسور بسيار حساس جهت رديابي آنتي بيوتيك كانامايسين
مقطع تحصيلي :
كارشناسي ارشد
گرايش تحصيلي :
بيوتكنولوژي
محل تحصيل :
اصفهان: دانشگاه صنعتي اصفهان، دانشكده كشاورزي
صفحه شمار :
دوازده، 58ص.: مصور، جدول، نمودار
يادداشت :
ص.ع. به فارسي و انگليسي
استاد راهنما :
مجيد طالبي
توصيفگر ها :
كانامايسين , آپتاسنسور , Real Time PCR , امنيت غذايي
استاد داور :
امير مساح، مهدي كديور
تاريخ ورود اطلاعات :
1396/07/02
چكيده انگليسي :
59 A Novel Ultrasensitive DNA Aptasensor for Kanamycin Detection Maedeh Setayesh Nasab Setayesh nasab@yahoo com August 30 2017 Department of Agricultural Biotechnology Isfahan University of Technology Isfahan 84156 83111 IranDegree M SC Language FarsiSupervisor M Talebi Ph D Assoc Professor mtalebi@cc iut ac irAbstractAntibiotics are widely used to eliminate or inhibit microorganisms Kanamycin Kn is one of theantibiotics used in animal feed to treat diseases and improve growth and its residues in animal products isa global concern Kanamycin residues in animal products can cause serious side effects such as loss ofhearing toxicity to the kidneys and allergic reactions to the drugs Therefore an accurate and easysystem is essential for detection of Kn The traditional methods used in food quality are time consuming difficult and require expensive equipment Therefore developing effective methods for food safety isneeded Aptamer based methods can overcome the disadvantages of traditional methods In this research a new type of biosensor based on an aptamer specific to Kn is developed The Kn aptamer is as amolecular detection probe and its complementary DNA acts as a signal generator for amplification inReal Time PCR In this study the results of five concentrations of 10 10 4 nM of complementary DNAwere investigated and the optimal concentration of 10 nM with minimum Ct was selected Afteroptimizing the concentration of complementary DNA three concentrations of 2 5 5 and 10 ng ml 1 ofstreptavidin and five concentration of 0 5 10 15 and 20 nM of aptamer were investigated and the lowestCt for aptamer and streptavidin was obtained at a concentration of 10 nM and 1 ng ml 1 respectively Also four concentrations of 0 2 5 5 and 10 ng ml 1 of streptavidin were studied and concentration of 5ng ml 1 with the lowest Ct was selected as the best concentration Under optimal conditions a lineardetection range standard curve for different concentrations of Kn 5 to 5X10 5 was obtained R2 0 991 and the limit of detection was 50 fgmL 1 in the range of 5 to 5X10 5 The specificity of this aptasensor wasexcellent compared with six other antibiotics Chloramphenicol Rifampicilin Cefotaxime Tetracycline Ampicilin and Streptomycin and perceptible Ct value change was not observed In addition the validityof this aptamer was appraised by identification of Kn spiked into water 0 5 5X10 2 and 5X10 4 ngmL 1 and milk 0 5 5X10 3 and 5X10 5 ngmL 1 and the recoveries in water and milk were in the range of 96 and 110 Therefore this method indicates an inexpensive very sensitive and specific for Kn detectionand can be used with great ability for detection of a wide range of target analytes in a single PCR tube Keywords Kanamycin Aptasensor Real Time PCR Food security
استاد راهنما :
مجيد طالبي
استاد داور :
امير مساح، مهدي كديور
