The Epideiologic Study And The Construction Of Detection DNA Microarray For Common Virues Of Shrimp

With the arrival of high-density cultural mode in shrimp, viral diseases have been causing more and more loss which has seriously restricted the development of aquaculture shrimp. The viruses of shrimp in culture zones exist widely in the body of crustaceans and have some complex routes of transmission. Combined with strenuous research of aquatic researchers in pathogenic biology research and demonstration breeding test, some valuable advice have taken shape such as asserted that establishing healthy breeding concepts, adopting comprehensive prevention measures on management and periodically pathogenic test for every stage of the shrimp to effectively prevent virus disease occurrence.This paper firstly conducted common virus disease epidemiological analysis of shrimp and demonstrated the prevalence trend of virus disease in shrimp. By conducting shrimp species and virus disease relations, breeding area and virus incidence relation, breeding season and the virus incidence relation and virus positive ratio analysis, we conclude that WSSV,IHHNV,HPV are the most three main pathogen that cause the virus disease. Compared to 34.90% of IHHNV and 11.98% of HPV, the rate of infection of WSSV of total virus infection has accounted for 50.52%.The existing technologies of pathogenic detection, however, are considered as less efficiency and poor systemic features, some strategic improvements are necessary for marine animal diseases high-throughput detection means. Along with the based information data of pathogenic of aquatic animal increasingly plentiful, trend of technology development in aquatic animal diseases diagnosis inevitably toward high-throughput technical direction across. During the 1990s the gene chip technology quickly developed with the advantages of high throughput and high efficiency in researching gene. It is significant to applying the new method to detect the pathogen from marine animals.According to the Blue Lightner, the Office International Des Epizooties (OIE) and the Network of Aquaculture Centers in Asia-Pacific (NACA), and other aquatic animals diseases directory, the research selected eight kinds of viruses, causing viral diseases in shrimp, as the research object and referenced to the previous study which provided 27 pairs of primer and 54 probes corresponding to the eight viruses in shrimp. Under this basis, we got a group of primers and probes which could be applied to multiplex RT-PCR and gene chip.In this study, We have attempted to develop a multiplex reverse transcription-polymerase chain reaction for simultaneously detection of six major shrimp viruses including white spot syndrome virus (WSSV), infectious hypodermal and hematopoietic necrosis virus (IHHNV), hepatopancreatic parvovirus (HPV), Taura syndrome virus (TSV), Baculovirus penae (BP) and infectious myonecrosis virus(IMNV). The reaction condition was optimized ,and the specificity and sensitivity was tested for this method.. The detection for real samples have also proved its reliability.Combined to the multiplex RT-PCR, we tried to found the technology used to detect eight viruses of shrimp. In this study, we have designed the DNA array for gene chip including positive quality control, negative quality control, external quality control and inner quality control and the hybrid area of probe of the corresponding viruses. Meanwhile, we have also tested the feasibility, specificity and applicability for gene chip.