Merino Victor R, Nakano Viviane, Delannoy Sabine, Fach Patrick, Alberca Gabriela G F, Farfan Mauricio J, Piazza Roxane M F, Avila-Campos Mario J
Laboratório de Anaeróbios, Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade of São Paulo, São Paulo, Brazil.
Agence Nationale de Sécurité Sanitaire de l'Alimentation, de l'Environnement et du Travail, Food Safety Laboratory, Université Paris-Est, Champs-sur-Marne, France.
Front Cell Infect Microbiol. 2020 Sep 25;10:549919. doi: 10.3389/fcimb.2020.549919. eCollection 2020.
The use of molecular diagnostics for pathogen detection in epidemiological studies have allowed us to get a wider view of the pathogens associated with diarrhea, but the presence of enteropathogens in asymptomatic individuals has raised several challenges in understanding the etiology of diarrhea, and the use of these platforms in clinical diagnosis as well. To characterize the presence of the most relevant bacterial enteropathogens in diarrheal episodes, we evaluated here the prevalence of diarrheagenic pathotypes, spp., and in stool samples of children with and without diarrhea using real-time quantitative PCR (qPCR). We found that the presence of genetic markers associated with bacterial pathogens was significantly higher in stool samples from the diarrhea group compared to the control ( < 0.001). Bacterial loads in samples positive for and markers were also determined. Compared to samples from asymptomatic children, a significantly higher number of copies of the gene were found in diarrhea samples. Also, the presence of genetic markers associated with STEC strains with clinical significance was evaluated in -positive samples by high-throughput real-time PCR. The data presented herein demonstrated that asymptomatic children of an urban area in Brazil might be enteropathogen reservoirs, especially for STEC.
在流行病学研究中使用分子诊断技术进行病原体检测，使我们能够更全面地了解与腹泻相关的病原体。然而，无症状个体中存在肠道病原体，这在理解腹泻病因以及将这些检测平台用于临床诊断方面都带来了诸多挑战。为了明确腹泻发作中最相关的细菌性肠道病原体的存在情况，我们在此使用实时定量聚合酶链反应（qPCR）评估了致泻性致病型、菌种以及在腹泻儿童和无腹泻儿童粪便样本中的流行情况。我们发现，与对照组相比，腹泻组粪便样本中与细菌病原体相关的基因标记物的存在显著更高（<0.001）。还测定了和标记物呈阳性的样本中的细菌载量。与无症状儿童的样本相比，腹泻样本中基因的拷贝数明显更多。此外，通过高通量实时PCR在呈阳性的样本中评估了与具有临床意义的产志贺毒素大肠杆菌（STEC）菌株相关的基因标记物的存在情况。本文呈现的数据表明，巴西一个城市地区的无症状儿童可能是肠道病原体的储存宿主，尤其是对于STEC而言。
