Halada P, Hlavackova K, Risueño J, Berriatua E, Volf P, Dvorak V
Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech Republic.
Department of Parasitology, Faculty of Science, Charles University, Prague, Czech Republic.
Med Vet Entomol. 2018 Sep;32(3):388-392. doi: 10.1111/mve.12305. Epub 2018 May 18.
Sandflies (Diptera: Psychodidae) (Newstead, 1911) are blood-feeding insects that transmit human pathogens including Leishmania (Trypanosomatida: Trypanosomatidae) parasites, causative agents of the leishmaniases. To elucidate Leishmania transmission cycles, conclusive identification of vector species is essential. Molecular approaches including matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) protein profiling have recently emerged to complement morphological identification. The aim of this study was to evaluate the effect of the trap type used to collect sandflies, specifically Centers for Disease Control (CDC) light or sticky traps, the two most commonly used in sandfly surveys, on subsequent MALDI-TOF MS protein profiling. Specimens of five species (Phlebotomus ariasi, Phlebotomus papatasi, Phlebotomus perniciosus, Phlebotomus sergenti, Sergentomyia minuta) collected in periurban and agricultural habitats in southeast Spain were subjected to protein profiling. Acquired protein spectra were queried against an in-house reference database and their quality assessed to evaluate the trap type effect. The results indicate that trap choice can substantially affect the quality of protein spectra in collected sandflies. Whereas specimens retrieved from light traps produced intense and reproducible spectra that allowed reliable species determination, profiles of specimens from sticky traps were compromised and often did not enable correct identification. Sticky traps should therefore not be used in surveys that deploy MALDI-TOF MS protein profiling for species identification.
白蛉（双翅目：蛾蠓科）（纽斯特德，1911年）是吸食血液的昆虫，可传播包括利什曼原虫（动基体目：锥虫科）寄生虫在内的人类病原体，这些寄生虫是利什曼病的病原体。为了阐明利什曼原虫的传播周期，对媒介物种进行确凿鉴定至关重要。包括基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）蛋白质谱分析在内的分子方法最近已出现，以补充形态学鉴定。本研究的目的是评估用于收集白蛉的诱捕器类型，特别是疾病控制中心（CDC）的灯光诱捕器或粘性诱捕器（这两种是白蛉调查中最常用的诱捕器）对后续MALDI-TOF MS蛋白质谱分析的影响。对在西班牙东南部城郊和农业栖息地采集的五个物种（阿氏白蛉、巴氏白蛉、中华白蛉、氏白蛉、微小赛蛉）的标本进行了蛋白质谱分析。将获得的蛋白质谱与内部参考数据库进行比对，并评估其质量以评估诱捕器类型的影响。结果表明，诱捕器的选择会极大地影响所收集白蛉蛋白质谱的质量。从灯光诱捕器中获取的标本产生的光谱强烈且可重复，能够可靠地鉴定物种，而从粘性诱捕器中获取的标本的谱图则受到影响，通常无法进行正确鉴定。因此，在采用MALDI-TOF MS蛋白质谱分析进行物种鉴定的调查中不应使用粘性诱捕器。
