Improvement of polymerase chain reaction-based Bt11 maize detection method by reduction of non-specific amplification.
نویسندگان
چکیده
The Bt11 maize-specific qualitative detection method based on polymerase chain reaction (PCR) in the JAS analytical test handbook has been widely used for administrative monitoring of GM crops and quality control of commercially distributed grains. In the present investigation, some apparently false-positive detections were observed in assays using the Bt11 maize-specific method, and these erroneous results were proved to have been caused by non-specific DNA amplification. We improved the detection method to reduce non-specific amplification by decreasing the concentration of magnesium ions in the PCR mixture. The subsequent evaluation of analytical performance demonstrated no marked difference between the currently used and the improved methods, except for the reduced non-specific amplification. We conclude that the currently used standard method should be replaced with the improved method for the reliable detection of Bt11 maize.
منابع مشابه
Polymerase chain reaction method for the rapid detection of virulent Shigella spp.
Bacillary dysentery, or shigellosis, is a disease of humans in which the colonic epithelium is invaded by bacteria and subjected to inflammatory destruction. The aim of this study was to develop a polymerase chain reaction(PCR) test for detection of virulent Shigella spp.. For this purpose, the primers were designed to amplify a 526-bp internal region of the Shigella spp. icsA gene, which encod...
متن کاملDevelopment of Multiplex Reverse Transcription-Polymerase Chain Reaction for Simultaneous Detection of Influenza A, B and Adenoviruses
Background and objective:Millions of people in developing countries lose their lives due to acute respiratory infections, such as Influenza A & B and Adeno viruses. Given the importance of rapid identification of the virus, in this study the researchers attempted to design a method that enables detection of influenza A, B, and adenoviruses, quickly and simultaneously. ...
متن کاملRAPID DETECTION OF MYCOBACTERIUM TUBERCULOSIS IN CLINICAL SPECIMENS BY POLYMERASE CHAIN REACTION
We investigated the use of DNA amplification by polymerase chain reaction (peR) for detection of Mycobacterium tuberculosis in 300 patients who were suspected of having pulmonary tuberculosis and compared the results with culture results which were performed in parallel with PCR. Two-thirds of each sample was processed for smear and culture by standard methods and one-third was prepared fo...
متن کاملEvaluation of Nucleic Acid Sequence Based Amplification (NASBA) and Reverse Transcription Polymerase Chain Reaction for Detection of Coxsackievirus B3 in Cell Culture and Animal Tissue Samples
Enteroviruses are the causative agents of a number of diseases in humans. Group B coxsackieviruses are believed to be the most common viral agents responsible for human heart disease. Genomic data of enteroviruses has allowed developing new molecular approaches such as Nucleic Acid Sequence Based Amplification (NASBA) for detection of such viruses. In this study, coxsackievirus B3 (CVB3) was de...
متن کاملSPECIFIC AMPLIFICATION OF ASPERGILLUS FUMlGATUS DNA BY POLYMERASE CHAIN REACTION
Invasive aspergillosis (1 is a life-threatening condition in immunocompromised patients. An early diagnosis is of great importance because early treatment may resolve this potentially fatal infection. Recently, the polymerase chain reaction (PCR) has been used successfully in detecting specific DNA of several pathogen. In this study, nested PCR was used to detect DNA specific for A!.pergiflus s...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Shokuhin eiseigaku zasshi. Journal of the Food Hygienic Society of Japan
دوره 51 1 شماره
صفحات -
تاریخ انتشار 2010