University of Florida, Institute of Food and Agricultural Sciences, Department of Entomology and Nematology, Florida Medical Entomology Laboratory, Vero Beach, Florida 32962, USA
University of Florida, Institute of Food and Agricultural Sciences, Department of Entomology and Nematology, Florida Medical Entomology Laboratory, Vero Beach, Florida 32962, USA.
Cold Spring Harb Protoc. 2024 Oct 1;2024(10):pdb.prot108291. doi: 10.1101/pdb.prot108291.
Mosquito species vary in their host associations. Although some species are relative generalists, most specialize, to varying extents, on particular types of host animals. Mosquito host associations are among the most important factors that influence the transmission dynamics of mosquito-vectored pathogens, and understanding these associations can provide insight on how such pathogens move within ecosystems. Characterization of the host associations of mosquito species requires applying blood meal analysis to the largest possible sample size of mosquito blood meals. Processing large samples of mosquito blood meals can be time-consuming, especially when chain-termination sequencing is used, necessitating individual processing of each specimen. Various methods and commercially available kits and products are available for extracting DNA from mosquito blood meals. The hot sodium hydroxide and Tris (HotSHOT) method is a rapid and inexpensive method of DNA extraction that is compatible with the recovery of DNA from mosquito blood meals preserved on QIAcard Flinders Technology Associates (FTA) Classic Cards (FTA cards). FTA cards allow nucleic acids found in blood meals to be preserved easily, even in field conditions. DNA prepared using this method is suitable for polymerase chain reaction (PCR)-based blood meal analysis.
蚊子种类在宿主关联性上存在差异。虽然有些种类相对较为普遍，但大多数种类在不同程度上专门针对特定类型的宿主动物。蚊子的宿主关联性是影响蚊子传播病原体动态的最重要因素之一，了解这些关联性可以深入了解此类病原体在生态系统中的传播方式。蚊子种类的宿主关联性特征需要对尽可能大的蚊子血餐样本量进行血液分析。处理大量的蚊子血餐样本可能很耗时，尤其是在使用链终止测序时，需要逐个处理每个标本。有各种方法和市售试剂盒和产品可用于从蚊子血餐中提取 DNA。热氢氧化钠和 Tris（HotSHOT）法是一种快速且廉价的 DNA 提取方法，与从保存在 QIAcard Flinders Technology Associates (FTA) Classic 卡（FTA 卡）上的蚊子血餐中回收 DNA 兼容。FTA 卡可轻松保存血餐中的核酸，即使在野外条件下也是如此。使用该方法制备的 DNA 适用于聚合酶链反应（PCR）为基础的血餐分析。
