Establishment of Fluorescence LAMP Detection System for Pathogen of Moko Disease

【Objective】The purpose of the study is to establish a real-time fluorescence qualitative detection method based on the LAMP technology and the specific conservative primers of the pathogen of Moko disease（Ralstonia solanacearum, race 2）. To solve the problems such as low specificity, low sensitivity and long time-consuming of qualitative detection of conventional PCR, an accurate and efficient qualitative detection within an hour can be achieved, which will provide technical support for the detection and quarantine of prevention and control of the disease.【Method】The sequence of gene target was determined and the Primer software was used to design two pairs of internal and external primers. The LAMP reaction system consisted of two pairs of primers, DNA polymerase with characteristics of strand displacement, template DNA, betaine, Mg 2SO4, reaction buffer, fluorescent dye etc. By using real-time fluorescence LAMP method, a lid-opening detection could be avoided after reaction and the sensitivity, specificity and repeatability of this method for the qualitative detection of sample DNA could be validated effectively.【Result】The designed primers could successfully amplify the target gene. The real-time fluorescence LAMP detection system had good sensitivity with specificity and strong repeatability for the pathogen of Moko disease, which was 10 times higher than that of ordinary PCR.【Conclusion】 The fluorescence LAMP detection system shows high specificity, high sensitivity, short time-consuming and low possibility of pollution for the detection of the pathogen of Moko disease.