Establishment And Application The Diagnose Methods Of Hemangioma Of Avian Leukosis Infecting Layer Chickens

Hemangioma of Avian leukosis is character of epidermization blood blister on chickens’skin and internal organ mainly caused by ALV.In recent years,hemangioma of Avian leukosis ocuurs severity in our country. It happens during laying period.the laying rate was lowed, death and eliminate rate can be20%,it has resulted in great enconmy loss to our poultry industry.Avian leukosis、Reticuloendotheliosis、Marek’s disease、 Chicken infectious anemia are four main immunity suppression diseases in poultry industry. They can induce respective pathological changes in chickens organs.And also induce subclinical infection in chikens.Causing immunity suppression and interfere with Newcastle disease and Avian Influenza vaccine immunifaction effect.In this study we established ALV A, B. J subgroups and MDV, REV, CAV diagnose methods to investigate the main reason inducing hemangioma.Diagnose typical phymatosis materials to investigate the incidence of the co-infection of immunity suppression disease.1. Establishment of Hemangioma of Avian leukosis diagnose methodsAccording to the published sequence of the ALV A, B、J subgroups standard strains of RAV-1, RAV-2, HPRS-103sequences,the genes of env labile region were amplified by PCR using synthetic oligonucleotide primers.The expected fragments of387bp,255bp,439bp were amplified and cloned into PMD18-T-Vector separately,Then inverted to Escherichia coli DH5a,cloned and identifyicated、sequenced.At the same time synthetized MDV, REV, CAV diagnose primers,used positive strains to optimize these primers conditions; In this study we establishment the PCR diagnose methods of ALV A、B、 J strains, it’s specific、 sensitivity;we also established diagnose methods of MDV, REV, CAV,this give a foundation to hemangioma of Avian leukosis diagnose.2. Isolation and gp85gene sequence analysis of hemangioma of Avian leukosis virusIn order to study the main cause of hemangioma in commercial layer chickens,14typical hemangiomatosis materials were isolated and identified by pathological anatomy,DF-1cell culture, IFA and PCR from typical hemangioma materials, discovered that10materials were caused by ALV-J,the IFA results were positive.Then synthesized primers of PJ3/PJ4.The gp85genes were amplified and sequenced.Sequence analysis revealed that:the10strains shared87.5%-96.8%nucleotide homology at home and abroad,The nucleotide homology was88.7%-76.4%between10stranis;The induced amino acids homology was78.0%～96.9%;evolution analysis showed that the isolation from guangxi had the lowest relationship with other strains,PD-108had the most relationship with HPRS-103,even the strains from the same rigion had the lower relationship.3. The established diagnose method clinic applicationApplication the established diagnose methods of ALV A, B, J strains and M DV, REV, CAV detected materials from40fall phymatosis fowl crowds,collected210materials of liver, spleen and serum,the results showed that:ALV was sever ity in layer chickens,ALV and MDV co-infection was17.5%, ALV-J was epidemi c in layer chickens.ALV+MDV+REV and REV+MDV+CAVthree immunity suppre ssion diseases co-infection was2.5%, ALV+MDV+REV+CAV four diseases co-inf ection was2.5%.Through investigation discovered immunity suppression disease c o-infection was widespread.