Study On Avian Viral Disease Using Universal Probe Real Time PCR

Avian viral disease is a kind of the most serious disease which threating poultry industry, for viruses is insensitivity to antibiotic and there is no effect in clinic,so the immunoprophylaxis is the best effect method , and disease monitoring is becoming a necessity component element. The key of controlling viral disease is to find viruses in early and forecast epidemic tendency, but viral disease displays atypical form recent years mostly, and routine method of detection cann't final diagnosis quickly, So the quick,accurate and highly efficient assay to detect viruses is needed for the Modern productionReal-time fluorescent PCR is a new technology of detecting nucleic acid,It is already greatestly applied to detect bacterium,virus and inherited disease.Basing on the principl of PCR ,this study created fluorescent real-time PCR detection of avian viral disease using universal probe real time PCR.This research is divided into the following three parts:Part one,Isolate and identify the Neweastle Disease virus.We plan to isoate and identify all relating vituses in this research to study the epidemiology of viruses in China in last srveral years, but we have too little time to isoate and identify NDV. We isoated and identified NDV by allantoic cavity culture, indirect immunofluorescrence assay (IFA), hemagglutination, hemagglutination inhibition,RT-PCR/PCR, gene-chip. We isolated 28 NDV field strains from 53 chickens which comed from 9 poultry industry of 7 province all over country, these results indicated ND was common in our country and gene-chip was a quick,accurate and highly efficient assay.Part two, The consructon of universal probe real time PCR detecting of Avian viral diseaseWe constructed a plasmid including a specified fragment of NDV, and optimizated reactive condition of universal probe real time PCR, and detected Infectious Laryngotracheitis Viruse, Infectious Bronchitis Viruse and...