目的 探索基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）方法用于蚊种鉴定的样本条件，为建立蚊种鉴定的MALDI-TOF MS方法提供依据，以快速、准确鉴定蚊虫种类。方法 以白纹伊蚊、淡色库蚊、三带喙库蚊和中华按蚊4种实验室品系蚊虫为研究对象，应用MALDI-TOF MS方法，对4种蚊虫的不同身体部位（整体、头部、胸部、足部）、2种保存方式（-20 ℃冷冻保存和硅胶干燥保存）以及2种吸血状态（未吸血和吸血）样本，采集肽质量指纹谱（PMF），建立蚊虫种类鉴定指纹谱数据库。同时用120个样本进行内部交叉验证，使用软件标准参数鉴定分值评价该数据库的准确性。结果 除腹部外，每种蚊虫的胸部、足部、头部指纹谱均可聚成一类，不同保存方法和饱血状态的蚊虫均未对蚊虫种类聚类结果产生影响。经数据库内部交叉验证，120个样本与形态学鉴定结果一致，鉴定分值均>2.00。1 μl的蚊虫蛋白提取液即可完成种类鉴定。结论 蚊虫的头、胸、足部样本均可用于MALDI-TOF MS法鉴定蚊虫种类，且蚊虫吸血状态和样本保存条件对蚊虫的鉴定结果无影响。MALDI-TOF MS可用于蚊虫种类鉴定，且具有快速、准确、灵敏等优点，对蚊种鉴定具有较高的使用价值。
葛文君, 肖迪, 张慧芳, 赵春春, 伦辛畅, 宋秀平, 郭玉红, 梁莹, 刘起勇, 孟凤霞
. 基于MALDI-TOF MS的蚊虫种类鉴定技术初步研究[J]. 中国媒介生物学及控制杂志, 2025
, 36(2)
: 238
-245
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DOI: 10.11853/j.issn.1003.8280.2025.02.016
Objective To explore the sample conditions for the matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF MS) method used for mosquito species identification, so as to provide a basis for establishing a MALDI-TOF MS method for rapid and accurate mosquito species identification. Methods Four laboratory mosquito strains, namely Aedes albopictus, Culex pipiens pallens, Cx. tritaeniorhynchus, and Anopheles sinensis, were selected as the study subjects. The MALDI-TOF MS method was used to analyze the samples from these mosquitoes for collecting peptide mass fingerprint and establishing a fingerprint database for mosquito species identification. The samples varied by body part (whole body, head, thorax, and foot), preservation method (cryopreservation at -20°C or dry preservation in silica gel), and blood-sucking status (non-blood-sucked and blood-sucked). The accuracy of this database was evaluated through internal cross-validation with 120 samples and identification scores using the software's standard parameters. Results Except for the abdomen, the fingerprints of the thorax, foot, and head of each mosquito species were clustered into one group, and different preservation method and blood-feeding status of those mosquitoes did not have any impact on the clustering results of mosquito species identification. At the same time, 120 samples were cross validated within the database, and the results were consistent with the morphological identification results, with identification scores >2.00. Only 1 μl of protein extract was allowed for the identification of mosquito species. Conclusions The head, thorax, and legs of mosquitoes can all be used for mosquito species identification by the MALDI-TOF MS method, and the blood-feeding status of mosquitoes and sample preservation methods do not affect mosquito identification results.The MALDI-TOF MS method can be used for mosquito species identification and offers the advantages of rapidity, accuracy, and sensitivity. These features make it highly useful for mosquito species identification.
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