A subgroup J avian leukosis virus (AVL-J), designated as ZH-08, was isolated from a breeder flock in Guangdong province with a novel hemangioma case. The identification results of ELISA test, PCR and immunofluoresecen...A subgroup J avian leukosis virus (AVL-J), designated as ZH-08, was isolated from a breeder flock in Guangdong province with a novel hemangioma case. The identification results of ELISA test, PCR and immunofluoresecence assay (IFA) specific for ALV-J were all positive. Based on the public full-length proviral genome sequence of ALV-J prototype strain HPRS-103, three pairs of primers were synthesized. The full-length proviral genome sequence of ZH-08 isolate is 7597 bp, which has a little difference with that of published full-length genome sequences, but its organization corresponds with typical retroviral genome structure; and known oncogenes were not included in its genome. According to the gp85 sequence comparison of ZH-08 isolate with those of the other reference strains in China and abroad, the highest similarity (93.7%) was with the YZ9901 isolate. Phylogenetic analysis, based on the gp85 gene, showed that the ZH-08 isolated here had the closest linkage to the SD07LK1 isolate. This study provides the basis for the biological characterization and pathogenesis research of the ZH-08 isolate.展开更多
An exogenous avian leukosis virus (ALV) strain SDAU09C1 was isolated in DF-1 cells from one of 240 imported 1-day-old white meat-type grand parent breeder chicks. Inoculation of SDAU09C1 in ALV-free chickens induced a...An exogenous avian leukosis virus (ALV) strain SDAU09C1 was isolated in DF-1 cells from one of 240 imported 1-day-old white meat-type grand parent breeder chicks. Inoculation of SDAU09C1 in ALV-free chickens induced antibody reactions specific to subgroup A or B. But gp85 amino acid sequence comparisons indicated that SDAU09C1 fell into subgroup A; it had homology of 88.8%-90.3% to 6 reference strains of subgroup A, much higher compared to other subgroups including subgroup B. This is the first report for ALV of subgroup A isolated from imported breeders.展开更多
Rapid diagnostic methods for classifying avian leukosis subgroups in the field were needed for routine, large-scale screening. As a first step in method development, we inserted the avian leukosis virus subgroup A （A...Rapid diagnostic methods for classifying avian leukosis subgroups in the field were needed for routine, large-scale screening. As a first step in method development, we inserted the avian leukosis virus subgroup A （ALV-A） env gene into plasmid pcDNA3.1/Zeo （＋） and used this construct to transfect DF-1 cells. Zeocin-resistant cells were obtained after 2 weeks of zeocin selection. Then, the cells were analyzed using PCR, immunofluorescence, and Western blot for expression of the envA-encoded envelope protein after 30 serial passages. The DF-1/A cell line was completely resistant to 104 TCIDso/0.1 mL （50% tissue culture infective dose）ALV-A and was partially resistant to 10~ TCIDs0/0.1 mL ALV-A viral particles. By comparing the DF-1/A and DF-1 cell lines, an ALV-A isolate was identified using a gag-specific ELISAfor capsid protein p27. Thus, we established a DF-1/A cell line that was resistant to ALV-A infection. This cell line will be useful as a diagnostic tool.展开更多
基金supported by National Science Foundation of Guangdong Province(Grant No.8151064201000065)Special Fund for Agro-scientific Research in the Public Interest(200803019)to Weisheng Cao+2 种基金NSFC-Guangdong Union Foundation(GrantNo.U0831002)National Natural Science Foundation(Grant No.30771612)Key Program of Science and Technology Development of Guangdong Province(Grant No.2009A020101006)to Ming Liao
文摘A subgroup J avian leukosis virus (AVL-J), designated as ZH-08, was isolated from a breeder flock in Guangdong province with a novel hemangioma case. The identification results of ELISA test, PCR and immunofluoresecence assay (IFA) specific for ALV-J were all positive. Based on the public full-length proviral genome sequence of ALV-J prototype strain HPRS-103, three pairs of primers were synthesized. The full-length proviral genome sequence of ZH-08 isolate is 7597 bp, which has a little difference with that of published full-length genome sequences, but its organization corresponds with typical retroviral genome structure; and known oncogenes were not included in its genome. According to the gp85 sequence comparison of ZH-08 isolate with those of the other reference strains in China and abroad, the highest similarity (93.7%) was with the YZ9901 isolate. Phylogenetic analysis, based on the gp85 gene, showed that the ZH-08 isolated here had the closest linkage to the SD07LK1 isolate. This study provides the basis for the biological characterization and pathogenesis research of the ZH-08 isolate.
基金supported by a grant from Shandong province and a grant (200803019) from the Ministry of Agriculturesupported by a grant from Shandong province and a grant (20080019) from the Ministry of Agriculture
文摘An exogenous avian leukosis virus (ALV) strain SDAU09C1 was isolated in DF-1 cells from one of 240 imported 1-day-old white meat-type grand parent breeder chicks. Inoculation of SDAU09C1 in ALV-free chickens induced antibody reactions specific to subgroup A or B. But gp85 amino acid sequence comparisons indicated that SDAU09C1 fell into subgroup A; it had homology of 88.8%-90.3% to 6 reference strains of subgroup A, much higher compared to other subgroups including subgroup B. This is the first report for ALV of subgroup A isolated from imported breeders.
基金The work was founded by the National Key R&D Program of China(2016YFD0501606)the Public Industry Research Program,the Ministry of Agriculture of China(201203055)+2 种基金the Program of Science and Technology Development of Guangdong Province,China(2015A020209145)the China Meat-Type Chicken Research System(CARS-42-G09)the Modern Agriculture Talents Support Program,Ministry of Agriculture of China([2012] no.160)
文摘Rapid diagnostic methods for classifying avian leukosis subgroups in the field were needed for routine, large-scale screening. As a first step in method development, we inserted the avian leukosis virus subgroup A （ALV-A） env gene into plasmid pcDNA3.1/Zeo （＋） and used this construct to transfect DF-1 cells. Zeocin-resistant cells were obtained after 2 weeks of zeocin selection. Then, the cells were analyzed using PCR, immunofluorescence, and Western blot for expression of the envA-encoded envelope protein after 30 serial passages. The DF-1/A cell line was completely resistant to 104 TCIDso/0.1 mL （50% tissue culture infective dose）ALV-A and was partially resistant to 10~ TCIDs0/0.1 mL ALV-A viral particles. By comparing the DF-1/A and DF-1 cell lines, an ALV-A isolate was identified using a gag-specific ELISAfor capsid protein p27. Thus, we established a DF-1/A cell line that was resistant to ALV-A infection. This cell line will be useful as a diagnostic tool.
