Berényi Olga, Bakonyi Tamás, Derakhshifar Irmgard, Köglberger Hemma, Topolska Grazyna, Ritter Wolfgang, Pechhacker Hermann, Nowotny Norbert
Zoonoses and Emerging Infections Group, Clinical Virology, Clinical Department of Diagnostic Imaging, Infectious Diseases and Clinical Pathology, University of Veterinary Medicine, Vienna, A-1210 Vienna, Austria.
Appl Environ Microbiol. 2007 Jun;73(11):3605-11. doi: 10.1128/AEM.00696-07. Epub 2007 Apr 13.
Honeybees originating from 10 different countries (Austria, Poland, Germany, Hungary, Slovenia, Nepal, Sri Lanka, the United Arab Emirates, Canada, and New Zealand) located on four continents were analyzed for the presence of deformed wing virus (DWV) nucleic acid by reverse transcription-PCR. Two target regions within the DWV genome were selected for PCR amplification and subsequent sequencing, i.e., a region within the putative VP2 and VP4 structural-protein genes and a region within the RNA helicase enzyme gene. DWV nucleic acid was amplified from 34 honeybee samples representing all the above-mentioned countries with the notable exception of New Zealand. The amplification products were sequenced, and phylogenetic analyses of both genomic regions were performed independently. The phylogenetic analyses included all sequences determined in this study as well as previously published DWV sequences and the sequences of two closely related viruses, Kakugo virus (KGV) and Varroa destructor virus 1 (VDV-1). In the sequenced regions, the DWV genome turned out to be highly conserved, independent of the geographic origins of the honeybee samples: the partial sequences exhibited 98 to 99% nucleotide sequence identity. Substitutions were most frequently observed at the same positions in the various DWV sequences. Due to the high level of sequence conservation, no significant clustering of the samples in the phylogenetic trees could be identified. On the other hand, the phylogenetic analyses support a genetic segregation of KGV and VDV-1 from DWV.
对来自四大洲10个不同国家(奥地利、波兰、德国、匈牙利、斯洛文尼亚、尼泊尔、斯里兰卡、阿联酋、加拿大和新西兰)的蜜蜂进行逆转录聚合酶链反应,以分析是否存在变形翅病毒(DWV)核酸。选择DWV基因组内的两个靶区域进行PCR扩增和后续测序,即假定的VP2和VP4结构蛋白基因内的一个区域以及RNA解旋酶基因内的一个区域。除新西兰外,从代表上述所有国家的34个蜜蜂样本中扩增出DWV核酸。对扩增产物进行测序,并对两个基因组区域分别进行系统发育分析。系统发育分析包括本研究确定的所有序列以及先前发表的DWV序列,以及两种密切相关病毒——角谷病毒(KGV)和瓦螨毁灭病毒1(VDV-1)的序列。在测序区域,DWV基因组高度保守,与蜜蜂样本的地理来源无关:部分序列显示出98%至99%的核苷酸序列同一性。在各种DWV序列的相同位置最常观察到替换。由于序列保守程度高,在系统发育树中无法识别出样本的明显聚类。另一方面,系统发育分析支持KGV和VDV-1与DWV的遗传分离。