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基于鸟枪法宏基因组学检测食品和水基质中的寄生虫:一篇叙述性综述。

Detection of parasites in food and water matrices by shotgun metagenomics: A narrative review.

作者信息

Vatta Paolo, Cacciò Simone M

机构信息

Unit of Foodborne and Neglected Parasitic Diseases, Department of Infectious Diseases, Istituto Superiore di Sanità, Viale Regina Elena 299, Rome 00161, Italy.

出版信息

Food Waterborne Parasitol. 2025 Apr 21;39:e00265. doi: 10.1016/j.fawpar.2025.e00265. eCollection 2025 Jun.

Abstract

Many helminths and protozoa are transmitted to humans through the consumption of contaminated food or water, and this underlines the importance of methods for their detection in these matrices. Due to the difficulties in isolating parasites prior to their identification, indirect detection methods are used, mostly relying upon targeted amplification of nucleic acids via PCR and/or qPCR. With the development of high throughput sequencing technologies, an untargeted detection method, shotgun metagenomics, became available. By sequencing the total DNA extracted from a given source, and through bioinformatics analyses of the sequencing reads, shotgun metagenomics allows profiling the entire microbial community therein present, including eukaryotes and, therefore, parasites. In this article, we reviewed the studies that specifically addressed the detection of parasites in food ( = 2) and water matrices ( = 10) by shotgun metagenomics. Most studies focused on wastewater samples and reported the detection of many parasites of human and veterinary importance from various areas of the world, highlighting the potential of shotgun metagenomics to provide important data for parasitic pathogens surveillance. After examining the different analytical workflows employed in these studies, which were not developed for detection of eukaryotes (or parasites), we identified two aspects deserving attention. First, that assignment based on short reads matching ribosomal sequences may generate false positives due to high sequence conservation among eukaryotic organisms. Second, that reassessing the relatively small number of reads of eukaryotic origin by a BLAST search can confirm, or deny, identification of parasitic pathogens.

摘要

许多蠕虫和原生动物通过食用受污染的食物或水传播给人类,这凸显了在这些基质中检测它们的方法的重要性。由于在鉴定之前分离寄生虫存在困难,因此使用间接检测方法,主要依靠通过PCR和/或qPCR对核酸进行靶向扩增。随着高通量测序技术的发展,一种非靶向检测方法——鸟枪法宏基因组学应运而生。通过对从给定来源提取的总DNA进行测序,并对测序读数进行生物信息学分析,鸟枪法宏基因组学能够对其中存在的整个微生物群落进行分析,包括真核生物,因此也包括寄生虫。在本文中,我们回顾了通过鸟枪法宏基因组学专门针对食品(n = 2)和水基质(n = 10)中寄生虫检测的研究。大多数研究集中在废水样本上,并报告了在世界不同地区检测到许多对人类和兽医具有重要意义的寄生虫,突出了鸟枪法宏基因组学为寄生虫病原体监测提供重要数据的潜力。在研究了这些并非为检测真核生物(或寄生虫)而开发的研究中采用的不同分析工作流程后,我们确定了两个值得关注的方面。第一,由于真核生物之间的高序列保守性,基于与核糖体序列匹配的短读数进行的分类可能会产生假阳性。第二,通过BLAST搜索重新评估相对较少的真核生物来源的读数可以确认或否定寄生虫病原体的鉴定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eaa9/12098152/7d6391a76574/ga1.jpg

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