Diagnostic Genechip for Diferentiation of Four Kinds of Vesicular Diseases

Abstract：Genechip technology was adopted to diferentiate the four animal vesicular diseases in order to distinguish them properly，rapidly an d largely．Thirty-nine amplified target gene fragments were puriried with Qiaquick 96 plate kit，and diluted with TE to obtain a concentration of 0．36 0．36／ag／／aL．A ． microarray was spotted with a GMS417 (pine．and．ring spotter)apparatus，after which the cDNA fragments were coupled with ．．NH2 on a genechip using ultraviolet cross linkage technique．Afterwards the quality of chip was confirmed by aspecific hybridization with salmon sperm ．Large quan tities of Foot．and．mouth disease virus(FMDV)RNA from infected tissues and Swine vesicular disease virus rSVDV)RNA from infected cell culture supematant were extracted and，fluoromarker were used to label cDNA fragments of FM DV an d SVDV when they am plified with random primers．After the labeled cDNA was randomly sheared by ultrasonic treatment，the broken pieces were SUbjected to specific hybridization，an d then photoscanned using optimal fight intensity，an d the result an alyzed with Imagene software．Comparison of the signals from the specimens while taking into consideration correspondent position of each virus on the genechip，allowed differential diagnoses of the four kinds of vesicular diseases．The diagnostic genechip method was rapid，sensitive and exact，rush through．