Plant Health and Environment Laboratory, Diagnostic and Surveillance Services, Ministry for Primary Industries, P.O. Box 2095, Auckland, 1140, New Zealand.
School of Biological Sciences, University of Auckland, Auckland, New Zealand.
Sci Rep. 2018 Jun 25;8(1):9623. doi: 10.1038/s41598-018-27603-x.
Small hive beetle (SHB), Aethina tumida can feed on honey, pollen and brood in honey bee colonies. It was endemic to Africa, but since 1996 has been detected in a number of countries worldwide, including Australia, Brazil, Canada, Italy, Mexico, South Korea, Philippines and the USA where it has had economic effects on local apiculture. To improve SHB identification, we obtained the first reference sequences from the DNA barcoding 5' COI gene region for SHB and some species of the family Nitidulidae associated with beehives. Phylogenetic analysis of SHB COI sequences (3' COI) revealed two divergent lineages, with those from Australia and USA being genetically different from the recent detection in Italy. Many countries, including New Zealand, are currently free from SHB, and require a rapid detection method for biosecurity. Here we present the development and validation of a real-time PCR assay for detection of SHB. The assay showed high specificity and sensitivity for detecting SHB, with no cross-reaction observed with closely related species, such as A. concolor. The real-time PCR is sensitive, detecting the target sequences up to 100 copies/µL. This assay should prove a useful biosecurity tool for rapid detection of SHB worldwide.
小蜂虻（SHB），Aethina tumida 可食用蜂蜜、花粉和蜜蜂幼虫。它原本是非洲特有的物种，但自 1996 年以来，已在全球多个国家被发现，包括澳大利亚、巴西、加拿大、意大利、墨西哥、韩国、菲律宾和美国，其对当地养蜂业造成了经济影响。为了提高 SHB 的识别能力，我们获得了 SHB 及一些与蜂巢有关的 Nitidulidae 科物种的 DNA 条形码 5' COI 基因区域的首个参考序列。对 SHB COI 序列（3' COI）的系统发育分析显示出两个分化的谱系，其中来自澳大利亚和美国的谱系在遗传上与最近在意大利的检测结果不同。包括新西兰在内的许多国家目前没有 SHB，需要一种快速的生物安全检测方法。在这里，我们提出了一种用于检测 SHB 的实时 PCR 检测方法的开发和验证。该检测方法对 SHB 具有高度的特异性和敏感性，与密切相关的物种（如 A. concolor）没有交叉反应。实时 PCR 检测方法灵敏度高，可检测到 100 拷贝/µL 的目标序列。该检测方法应该成为全球快速检测 SHB 的有用的生物安全工具。
