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基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)作为成年物种诊断工具的应用

The Use of MALDI-TOF MS as a Diagnostic Tool for Adult Species.

作者信息

Rivero Julia, Zurita Antonio, Cutillas Cristina, Callejón Rocío

机构信息

Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Seville, Seville, Spain.

出版信息

Front Vet Sci. 2022 May 11;9:867919. doi: 10.3389/fvets.2022.867919. eCollection 2022.

Abstract

Trichuriasis is considered a neglected tropical disease, being the second most common helminthiasis in humans. Detection of in routine diagnosis is usually done by microscopic detection of eggs in fecal samples. Other molecular analyses are more reliable and could be used, but these analyses are not routinely available in clinical microbiology laboratories. The use of matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) is increasing since the last decades due to its recent evidence as a potential role for reliable identification of microorganisms and a few nematodes. But, for parasites detection, normalized protocols and the acquisition and introduction of new species to the database are required. We carried out a preliminary study confirming the usefulness of MALDI-TOF MS for the rapid and reliable identification of used as control and the creation of an internal database. To create main spectra profiles (MSPs), the different parts of five whipworms (esophagus and intestine) were used, developing different tests to verify the repeatability and reproducibility of the spectra. Thus, to validate the new internal database, 20 whipworms, separating the esophagus and intestine, were used, of which 100% were accurately identified as , but could not distinguish between both parts of the worm. Log score values ranged between 1.84 and 2.36, meaning a high-quality identification. The results confirmed that MALDI-TOF MS was able to identify species. Additionally, a MALDI-TOF MS profile of proteome was carried out to develop the first internal database of spectra for the diagnosis of trichuriasis and other spp.

摘要

鞭虫病被认为是一种被忽视的热带疾病,是人类第二常见的蠕虫病。常规诊断通常通过显微镜检测粪便样本中的虫卵来进行。其他分子分析更可靠,可以使用,但这些分析在临床微生物实验室中并非常规可用。自过去几十年以来,基质辅助激光解吸/电离飞行时间(MALDI-TOF)质谱(MS)的使用不断增加,因为其最近被证明在可靠鉴定微生物和一些线虫方面具有潜在作用。但是,对于寄生虫检测,需要标准化方案以及将新物种纳入数据库。我们进行了一项初步研究,证实了MALDI-TOF MS在快速可靠鉴定用作对照的鞭虫以及创建内部数据库方面的有用性。为了创建主要光谱图谱(MSP),使用了五条鞭虫的不同部位(食管和肠道),开展了不同测试以验证光谱的可重复性和再现性。因此,为了验证新的内部数据库,使用了20条鞭虫,将食管和肠道分开,其中100%被准确鉴定为鞭虫,但无法区分虫体的两个部位。对数得分值在1.84至2.36之间,意味着高质量鉴定。结果证实MALDI-TOF MS能够鉴定鞭虫物种。此外,还进行了鞭虫蛋白质组的MALDI-TOF MS分析,以建立用于诊断鞭虫病和其他鞭虫属物种的首个内部光谱数据库。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf1e/9132177/f9d13d6780f7/fvets-09-867919-g0001.jpg

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