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评估一种多重聚合酶链反应检测法用于同时检测儿科患者粪便样本中的细菌和病毒肠道病原体。

Evaluation of a multiplex PCR assay for simultaneous detection of bacterial and viral enteropathogens in stool samples of paediatric patients.

作者信息

Onori Manuela, Coltella Luana, Mancinelli Livia, Argentieri Marta, Menichella Donato, Villani Alberto, Grandin Annalisa, Valentini Diletta, Raponi Massimiliano, Russo Cristina

机构信息

Department of Laboratory Medicine, Virology Unit, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy.

Department of Laboratory Medicine, Virology Unit, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy.

出版信息

Diagn Microbiol Infect Dis. 2014 Jun;79(2):149-54. doi: 10.1016/j.diagmicrobio.2014.02.004. Epub 2014 Feb 22.

Abstract

We evaluated a multiplex PCR assay, the Seeplex Diarrhoea ACE detection, that simultaneously detects 15 enteric pathogens, including Salmonella spp., Shigella spp., Vibrio spp., toxin B producer Clostridium difficile, Campylobacter spp., Clostridium perfringens, Yersinia enterocolitica, Aeromonas spp., Escherichia coli O157:H7, verocytotoxin-producing Escherichia coli, adenovirus, Group A rotavirus, norovirus GI and GII, and astrovirus. We compared this assay with clinical methods routinely used in our laboratory, for detecting enteropathogens in stool samples collected from 245 paediatric patients with suspected infectious gastroenteritis. We recovered 61 bacterial pathogens and 121 enteric viruses with our laboratory assays, while we detected 78 bacteria and 167 viruses with the molecular assay. We calculated specificity and sensitivity for both methods after analysis of discordant results and demonstrated greater sensitivity for multiplex PCR than for our routine methods, with the exception of Salmonella spp. and toxigenic C. difficile detection. The multiplex PCR assay proved to be a reliable tool to directly detect the most common enteropathogens in stool samples but with some limitations.

摘要

我们评估了一种多重聚合酶链反应检测方法——Seeplex腹泻ACE检测法,该方法可同时检测15种肠道病原体,包括沙门氏菌属、志贺氏菌属、弧菌属、产毒素B的艰难梭菌、弯曲杆菌属、产气荚膜梭菌、小肠结肠炎耶尔森菌、气单胞菌属、大肠杆菌O157:H7、产志贺毒素大肠杆菌、腺病毒、A组轮状病毒、诺如病毒GI和GII以及星状病毒。我们将该检测方法与我们实验室常规使用的临床方法进行了比较,以检测从245名疑似感染性肠胃炎的儿科患者收集的粪便样本中的肠道病原体。通过我们的实验室检测方法,我们检出了61种细菌病原体和121种肠道病毒,而通过分子检测方法,我们检测到了78种细菌和167种病毒。在对不一致结果进行分析后,我们计算了两种方法的特异性和敏感性,结果表明,除了沙门氏菌属和产毒素艰难梭菌的检测外,多重聚合酶链反应检测方法比我们的常规方法具有更高的敏感性。多重聚合酶链反应检测方法被证明是一种直接检测粪便样本中最常见肠道病原体的可靠工具,但也存在一些局限性。

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