Huang Wei-Fone, Zhang Yakun, Mehmood Shahid, Wang Zhengwei, Hou Chunsheng, Li Zhiguo
College of Animal Science (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Chemical Ecology Group, Key Laboratory of Tropical Forest Ecology, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Kunming 650000, China.
Vet Sci. 2021 Apr 13;8(4):63. doi: 10.3390/vetsci8040063.
Sacbrood virus (SBV) is a common honey bee virus disease. SBV variants and strains identified in Asian honey bees, , have created confusion in identifications. Although the regional names indicated the expansions of the virus in new regions, pathogenesis, and genomes of these variants are not distinct enough to be a separate virus species. However, current SBV qPCR methods may not detect newly identified SBV variants (Ac SBV) according to the genome sequences. Since these Ac SBV can naturally infect and possibly other hymenopterans, ignorance of Ac SBV variants in detection methods is simply unwise. In this report, we updated the qPCR method based on Blanchard's design that used conserved regions of VP1 to design a TaqMan method with an MGB (minor groove binder) probe. We tested the method in bees and hornets, including , , and . The updated primers and the probe can match published SBV and Ac SBV genomes in databases, and this updated method has reasonable sensitivity and flexibility to be applied as a detection and quantification method before the discovery of variants with more mutated VP1 gene.
囊状幼虫病毒（SBV）是一种常见的蜜蜂病毒性疾病。在亚洲蜜蜂中鉴定出的SBV变体和毒株在鉴定方面造成了混乱。尽管这些区域名称表明该病毒在新区域有所传播，但其变体的发病机制和基因组差异并不足以构成一个单独的病毒物种。然而，根据基因组序列，目前的SBV定量聚合酶链反应（qPCR）方法可能无法检测到新鉴定出的SBV变体（Ac SBV）。由于这些Ac SBV能够自然感染，并且可能感染其他膜翅目昆虫，因此在检测方法中忽视Ac SBV变体是不明智的。在本报告中，我们基于布兰查德的设计更新了qPCR方法，该设计利用VP1的保守区域设计了一种带有小沟结合物（MGB）探针的TaqMan方法。我们在蜜蜂和黄蜂中测试了该方法，包括、和。更新后的引物和探针能够与数据库中已公布的SBV和Ac SBV基因组匹配，并且在发现具有更多VP1基因突变的变体之前，这种更新后的方法具有合理的灵敏度和灵活性，可作为一种检测和定量方法应用。
