Study On The Liquid Gene Chip Deteciton Technique To Shrimp Yellow Head Disease

Shrimp yellow head disease is a kind of shrimp infectious disease which causedby the yellow head virus (YHV). The appearance of dying shrimp's head and chest isyellow due to hepatopancreas become yellow, so it is called yellow head disease.Yellow head virus (YHV) belongs to the virales Nidovirales, family Ronivi-ridae,genus Okavirus. The disease is classified as II animal disease in China, and thedisease must be declared by the required of International office of Epizootic Disease(OIE).The disease was first reported in early1992, black tiger shrimp that were farmedin Nakornsrithammarant Pakpaneng which in southern Thailand infected yellow headdisease. In May2005, the first detection of yellow head virus from the two batches ofshrimp imported from Thailand was declared by Shanghai Entry-Exit Inspection andQuarantine Bureau. The threaten to Chinese shrimp farming industry is more andmore serious. It include two parts in this study:(1) The first step include acquisition of the pathological material and extractionof the viral RNA. RT-PCR experiments produce the cDNA of perfect length. A pair ofspecific primer was designed for amplifying the fragment in RT-PCR. Optimizing ofthe annealing temperature, Mg2+concentration, cycle number and other parameters ofthe PCR reaction is in order to get the needed DNA fragments.(2) According to the acquisition of experiment, two efficient fluorescent andcoded microspheres were selected. Microspheres and nucleic acid probe were coupledby EDC one-step. The specificity test showed that the coupling efficiency was veryhigh. The liquid cDNA chip analysis system of shrimp yellow head disease wasestablished and detect the gene of GP116successfully. Further optimization of thevarious conditions of the system is in order to detects shrimp Yellow Head Virus moreeasily.