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针对人类和动物皮肤癣菌病的皮肤癣菌的快速鉴定和新型蛋白质组学分析。

Towards a rapid identification and a novel proteomic analysis for dermatophytes from human and animal dermatophytosis.

机构信息

Department of Microbiology, Faculty of Veterinary Medicine, Zagazig University, Zagazig, Egypt.

Department of Bacteriology, Immunology and Mycology, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, Egypt.

出版信息

Mycoses. 2019 Dec;62(12):1116-1126. doi: 10.1111/myc.12998. Epub 2019 Oct 16.

Abstract

BACKGROUND

Since accurate identification of dermatophyte species is essential for epidemiological studies and implementing antifungal treatment, overcoming limitations of conventional diagnostics is a fruitful subject.

OBJECTIVES AND METHODS

In this study, we investigated real-time polymerase chain reaction(q-PCR), matrix-assisted laser desorption/ionisation time of flight mass spectrometry (MALDI-TOF MS) and nano-electrospray ionisation mass spectrometry (nano-ESI-MS) to detect and identify the most frequently isolated dermatophytes from human and animal dermatophytosis in comparison with conventional methods.

RESULTS

Among 200 samples, the identified species were Microsporum canis (78.22%), Trichophyton verrucosum (10.89%) and T. mentagrophytes (5.94%). Q-PCR assay displayed great execution attributes for dermatophytes detection and identification. Using MALDI-TOF MS, M. canis, but none of T. violacium, T. verrucosum or T. mentagrophytes, could be identified. Nano-ESI-MS accurately identified all species. The potential virulence attributes of secreted proteases were anticipated and compared between species. Secreted endoproteases belonging to families/subfamilies of metalloproteases, subtilisins and aspartic protease were detected. The analysed exoproteases are aminopeptidases, dipeptidyl peptidases and carboxypeptidases. Microsporum canis have three immunogenic proteins, siderophore iron transporter mirB, protease inhibitors, plasma membrane proteolipid 3 and annexin.

CONCLUSION

In essence, q-PCR, MALDI-TOF MS and nano-ESI-MS assays are very nearly defeating difficulties of dermatophytes detection and identification, thereby, supplement or supplant conventional diagnosis of dermatophytosis.

摘要

背景

由于准确识别皮肤癣菌对于流行病学研究和实施抗真菌治疗至关重要,因此克服传统诊断方法的局限性是一个富有成效的课题。

目的和方法

在这项研究中,我们调查了实时聚合酶链反应(q-PCR)、基质辅助激光解吸/电离飞行时间质谱 (MALDI-TOF MS) 和纳升电喷雾电离质谱 (nano-ESI-MS),以与传统方法相比,检测和鉴定人类和动物皮肤癣菌病中最常分离到的皮肤癣菌。

结果

在 200 个样本中,鉴定出的物种为犬小孢子菌(78.22%)、疣状毛癣菌(10.89%)和须癣毛癣菌(5.94%)。q-PCR 检测方法对皮肤癣菌的检测和鉴定具有出色的执行特性。使用 MALDI-TOF MS,只能鉴定出犬小孢子菌,而不能鉴定出 T. violacium、T. verrucosum 或 T. mentagrophytes。nano-ESI-MS 能够准确地鉴定所有物种。预测并比较了物种之间分泌蛋白酶的潜在毒力特性。检测到属于金属蛋白酶、枯草蛋白酶和天冬氨酸蛋白酶家族/亚家族的分泌内肽酶。分析的外肽酶是氨肽酶、二肽基肽酶和羧肽酶。犬小孢子菌有三种免疫原性蛋白,即铁载体 mirB、蛋白酶抑制剂、质膜疏脂蛋白 3 和膜联蛋白。

结论

本质上,q-PCR、MALDI-TOF MS 和 nano-ESI-MS 检测方法几乎克服了皮肤癣菌检测和鉴定的困难,从而补充或替代了皮肤癣菌病的传统诊断。

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