Research And Application Of Classical Swine Fever Virus Diagnostic Technology By Gene Chip

CSF(Classical swine fever), the virulent infectious disease caused by CSFV (Classical swine fever virus), had been controlled effectively by CSFV strain vaccine in the country. But atypical HC and inapparent CSF began to appear after 1980s, which had resulted in great loss. This result may have something to do with the appearance of the new variation virus of CSFV and other factors. Even sub-clinic infections swine is difficult to diagnose and eliminate them, depending on the general diagnosis method and bring new problem to the prevention and treatment. Thus it is necessary to establish a new, swift and reliable diagnosis system in the lab to pave a way for completely annihilating CSF.Gene chip or microarray, the combination of biology and microelectronics technology, is a kind of new micro-device. It is composed of DNA arrays immobilized on solid supports. Because of its many virtues, such as rapid, sensitive and accurate, gene chip plays an important role in the study of diseases. Through the match of mutation probes and sequences, we can detect the mutation quickly, lightly and accurately. The detection can effectively instruct disease diagnosis.The study utilized phylogenetic tree based on gene sequence to isolate and identify CSFVs in some areas of Heilongjiang province and sequence E2 gene of every virus. The results showed in the following: Doubtful materials were infected with CSFVs, the sequences of E2 genes nearer to Alfort, the homology with Brescia was the lowest. We researched the homology and variation among prevalent CSFVs on nucleic acid and protein level in some areas in the province. At the same time, the antigenic variation between traditional Shimen virus and virus cultivated in rabbits was compared in order to disclose the prevalent essence of CSF in the province and tried to acquire new efficient methods for preventing and treating CSF better and better.In this study a microarray-based assay for the detection CSFV was developed. The captor-DNA fragments, which were reamplified from CSFV C stain, were spotted on a glass-bound microarray. Cy5-labeled fluorescent PCR production were hybridized to these captor-DNA on microarray. At the same time, we studied the effects of the different concentration of the captor-DNA on the detecting result, confirmed the concentration of captor-DNA is 1μg/μl, established the option reaction condition, analyzed the specialty and sensitivity and compared it with RT-PCR and IFA experiment method. The result show that microarray diagnosis method of CSFV established in this research can successfullyverify the conservative sequence of the CSFV. The signal of the fluorescence is clear, the specialty is high.In fact it is an efficient diagnosis method.To sum up, this study successfully established a diagnosis technique platform of microarray and solved the difficult problem for CSFV, with afford a theoretical base for prevention and diagnosis of animal's epidemic diseases, control and annihilate CSF.But in order to abroad applicate microarray in animal disease,we need to do more reseach.