Development Of A GeXP Based Multiplex RT-PCR For Detection Of Potato Virus

Objective:It is expected to set a method how to check potato in a brand new and fast way, basing on the reaction of Genetic Analysis System assay method. This research was done with the technique of GeXP system, using the way of multiplex reverse transcription-polymerase chain reaction, testing7kinds of common virus and one virus-like agent of potato. Evaluate the result of this research.Methods:Set eight kinds of particular primers for eight kinds of potato viruses to organize a detection system which was suitable for multiple RT-PCT, and set eight potato viruses by the way of GeXP system. Improve the detection system and the condition of reaction in the process of testing. Prepare the PNA samples which were made by the method of in vitro transcription, testing the sensitivity with multiple RT-PCT detention system.Results:Through the screening of various virus primers, this research amplified fragment size consistent with design and strong specificity, and it had no cross reaction between each primer. After the improvement of multiple testing system and the optimized reaction conditions, it can amplify the specificity of corresponding segments at the same time. Meanwhile, it reached a high number of103copies/uL under a sensitivity. Through the test for100kinds of samples, it can be seen that GeXP multiple detection method specificity was strong and sensitiveConclusion:This research has successfully established a new and fast method to test potato virus. This method treats the technique of GeXP multiple detection method for reverse transcription RNA as a platform, compared with the traditional and simpler PCR, it has a higher coherence, sensitivity and speed, etc. It has a great significance of the investigation, selection, analysis and research about potato. And, it provides a new way of thinking gene test under a high-flux and multi-targets.