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Virologica Sinica, 18 (2) : 174-177, 2003
Diagnostic Genechip for Diferentiation of Four Kinds of Vesicular Diseases
Shenzhen Entry—exit Inspection and Quarantine Bureau,Shenzhen,5 18010,China
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
  Published online: 15 Apr 2003
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