Department of Biology, Case Western Reserve University, Cleveland, Ohio.
The Center for Global Health and Diseases, Case Western Reserve University, Cleveland, Ohio.
Am J Trop Med Hyg. 2019 Mar;100(3):544-551. doi: 10.4269/ajtmh.18-0782.
mosquitoes vary in habitat preference, feeding pattern, and susceptibility to various measures of vector control. Consequently, it is important that we identify reservoirs of disease, identify vectors, and characterize feeding patterns to effectively implement targeted control measures. Using 467 anopheline mosquito abdomen squashes captured in Madagascar, we designed a novel ligase detection reaction and fluorescent microsphere assay, dubbed Bloodmeal Detection Assay for Regional Transmission (BLOODART), to query the bloodmeal content, identify five mosquito species, and detect infection. Validation of mammalian bloodspots was achieved by preparation and analysis of known hosts (singular and mixed), sensitivity to degradation and storage method were assessed through mosquito feeding experiments, and quantification was explored by altering ratios of two mammal hosts. BLOODART identifications were validated by comparison with mosquito samples identified by sequenced portions of the internal transcribed spacer 2. BLOODART identification of control mammal bloodspots was 100% concordant for singular and mixed mammalian blood. BLOODART was able to detect hosts up to 42 hours after digestion when mosquito samples were stored in ethanol. A mammalian host was identified in every field-collected, blood-fed female mosquito by BLOODART. The predominant mosquito host was cow ( = 451), followed by pig ( = 26) and human ( = 25). Mixed species bloodmeals were commonly observed ( = 33). A BLOODART molecular identification was successful for 318/467 mosquitoes, with an overall concordance of 60% with all field-captured, morphologically identified specimens. BLOODART enables characterization of large samples and simultaneous pathogen detection to monitor and incriminate disease vectors in Madagascar.
蚊子在栖息地偏好、摄食模式和对各种病媒控制措施的敏感性方面存在差异。因此，确定疾病储存库、鉴定病媒、描述摄食模式对于有效实施有针对性的控制措施非常重要。我们使用在马达加斯加捕获的 467 个按蚊腹部压扁物，设计了一种新的连接酶检测反应和荧光微球检测法，称为区域传播血液检测分析（Bloodmeal Detection Assay for Regional Transmission，BLOODART），以查询血液内容物、鉴定 5 种蚊子种类并检测 感染。通过制备和分析已知宿主（单一和混合）来验证哺乳动物血斑，通过蚊子喂养实验评估对降解和储存方法的敏感性，并通过改变两种哺乳动物宿主的比例来探索定量分析。通过与内部转录间隔区 2 测序部分鉴定的蚊子样本进行比较来验证 BLOODART 鉴定。BLOODART 对单一和混合哺乳动物血液的控制血斑鉴定的一致性为 100%。当蚊子样本储存在乙醇中时，BLOODART 可以在消化后 42 小时内检测到宿主。BLOODART 可以在每只野外采集的、吸血的雌性 蚊子中鉴定出哺乳动物宿主。主要的蚊子宿主是牛（=451），其次是猪（=26）和人（=25）。常见的是混合物种的血餐（=33）。BLOODART 对 318/467 只蚊子进行了分子鉴定，与所有野外捕获的形态学鉴定的 标本总体一致性为 60%。BLOODART 能够对大量样本进行特征描述，并同时进行病原体检测，以监测和指控马达加斯加的病媒。
