Study On Part Of Microsatellite Instability In The Host Genome Of Marek’s Preneoplasia

Marek’s Disease is a common poultry communicable diseases. In addition to a report by the dynamic pathobiology study showed that Marek’s disease virus cells as well atherosclerosis as a basis for early stage diagnosis, there are no reports in molecular biology on the early stage diagnosis. It is extremely rare to using microsatellite instability to analyze reports of Marek’s disease tumors (in addition to the our laboratory) at home and abroad. In this trial, SPF white leghorn chickens were inoculated with HVT vaccine and infected wih MDV-RBIB strain, comprehensive observed and MSI in host genome was analyzed, to the discovery differences microsatellite sites showing MSI changes between HVT vaccine group and MDV-RBIB strain group. There are differences in the use of these microsatellite sites for early diagnosis provide test data.Twenty and40SPF chickens were inoculated with HVT vaccine and infected with MDV-RB1B strain differently, anticoagulated blood in wing vein was collected on a succession time; dissevered and tumor tissue, healthy muscle tissue was collected at the end of the experiment; the DNA of the blood lymphocytes, tumor tissue and healthy muscle tissue was extracted;46microsatellite markers were screened which showed MSI more than30%early in our laboratory infected MDV chicken embryo fibroblast cells of all the chromosomes in the chicken; MSI was tested by PCR amplification with the46microsatellite markers were designed46pairs primes, denaturing polyacrylamide gel electrophoresis and silver staining were performed.After SPF chickens were inoculated with HVT vaccine, MSI were tested by46microsatellite sites, the result showed that MSI changes of Peripheral blood lymphocytes happened on26microsatellite sites on the chickens, accounting for56.5%of the selected sites; in3days after immunizated, there were20microsatellite sites appering MSI, accounting for76.9%of the total changed sites; there were9MSI disappeared, accounting for34.6%of the changed sites. However, after SPF chickens were infected with MDV-RB1B strain, MSI were tested by46microsatellite sites which were the same to HVT vaccine guoup, the result showed that MSI changes of Peripheral blood lymphocytes and tumor tissue happened on30microsatellite sites on the infected chickens, accounting for65.2%of the selected sites. On the SPF chickens inoculated with MDV-RB1B, MSI changes of15microsatellite locis showed in tumor tissue and in peripheral blood lymphocytes had consistency which were ABR0365, MCW0145, ABR0262, ABR0086, ADL0147, ABR0659, ABR0052, ABR0352, ABR0634, ABR0389, LEI0099, ABR0007, ABR0617, ABR0107and ABR0387, accouting for50%of the total changed sites; and MSI changed of7microsatellite sites in tumor tissue and in peripheral blood lymphocytes had consistency but not showed in the peripheral blood lymphocyte on the HVT vaccine group, which were ABR0365, ABR0387, ABR0007, ABR0389, ABR0617, ABR0659and ADL0147.In this trial, SPF white leghorn chickens were inoculated with HVT vaccine and infected with MDV-RB1B strain differently, were observation and MSI in host genome were analyzed, the result showed that MSI changes of7microsatellite sites which are ABR0365、ABR0387、ABR0007、ABR0389、ABR0617、ABR0659and ADL0147have consistency in Peripheral blood lymphocytes and tumor tissues in Marek’s preneoplasia, but not showed in HVT vaccine group; ABR0033and ABR0399have MSI changed in Peripheral blood lymphocytes in Marek’s preneoplasia, but not showed in HVT vaccine group; MCW0145has MSI changed in4chicken’s Peripheral blood lymphocytes in Marek’s preneoplasia, but showed only1chick in HVT vaccine group. In other words, the13microsatellite sites which are ABR0365, ABR0033and MCW0145et al would be used to diagnose MD in Marek’s preneoplasia.