Zhang Zhi, Cui Zhizhong
Department of Preventive Veterinary Medicine, College of Animal Science and Veterinary Medicine, Shandong Agriculture University, Taian 271018, China.
Sci China C Life Sci. 2005 Feb;48(1):81-8. doi: 10.1360/03yc0270.
Two Marek's disease virus (MDV) field strains were isolated from chickens with tumors independently from Guangdong and Guangxi provinces, and it was confirmed that there were no co-infections with reticuloendotheliosis viruses (REV) in chicken embryo fibroblast cells (CEF) in indirect fluorescence antibody test (IFA) with REV-specific monoclonal antibodies. By dot blot hybridization and PCR of genomic DNA of MDV-infected CEF, it was indicated that LTR fragments of REV genome were integrated into genome of these two MDV field strains. To amplify and clone the integrated REV LTR with MDV sequence at the junction, 4 primers from REV LTR and 7 primers from MDV genome fragment with REV LTR insertion hot points were synthesized and 28 (4x7) pairs of primers (one from REV and another from MDV for each pair) were used in PCR while using the genomic DNA of both strains as the templates. The sequence data demonstrated that both recombinant field strains contained the same REV LTR inserted into MDV at the identical sites in US fragment of the genomes. From the above, it was speculated that both recombinant field MDVs were originated from a same recombinant virus and spread among chicken flocks in two provinces.
从广东省和广西壮族自治区独立采集的患有肿瘤的鸡中分离出两株马立克氏病病毒（MDV）野毒株，并且通过用网状内皮组织增生症病毒（REV）特异性单克隆抗体进行间接荧光抗体试验（IFA），证实在鸡胚成纤维细胞（CEF）中不存在与REV的共感染。通过对MDV感染的CEF的基因组DNA进行斑点杂交和PCR，表明REV基因组的长末端重复序列（LTR）片段整合到了这两株MDV野毒株的基因组中。为了扩增并克隆在连接处带有MDV序列的整合REV LTR，合成了4条来自REV LTR的引物和7条来自带有REV LTR插入热点的MDV基因组片段的引物，并且在以两株毒株的基因组DNA作为模板进行PCR时使用了28（4×7）对引物（每对引物中一条来自REV，另一条来自MDV）。序列数据表明，两株重组野毒株在基因组的US片段的相同位点均含有插入到MDV中的相同REV LTR。由此推测，两株重组野毒株MDV均起源于同一重组病毒，并在两个省份的鸡群中传播。
