Siegert Lisa, Stoll Sylvia, Edel Birgit, Löffler Bettina, Rödel Jürgen
Institute of Medical Microbiology, Jena University Hospital, Friedrich Schiller University of Jena, Jena, Germany.
Institute of Medical Microbiology, Jena University Hospital, Friedrich Schiller University of Jena, Jena, Germany.
Diagn Microbiol Infect Dis. 2025 Nov;113(3):117004. doi: 10.1016/j.diagmicrobio.2025.117004. Epub 2025 Jul 10.
Molecular assays, which are commonly based on multiplex PCR techniques, are increasingly being used to diagnose bacterial gastroenteritis due to faster results and more straightforward workflows compared to conventional culture. Many culture-dependent methods and semi-automated PCR assays, are labour-intensive and require technical expertise. Therefore, assays that are easy to perform and allow for the timely identification of the most common enteric bacterial pathogens may be of interest. This study investigated a molecular assay based on loop-mediated isothermal amplification (LAMP) for the rapid identification of several common bacterial pathogens. A total of 204 stool samples were analysed. The sensitivity and specificity of the assay, compared to the BD MAX™ Enteric Bacterial Panel PCR as the reference method, were as follows: 88.35 % and 99.04 % for Campylobacter spp., 88 % and 100 % for Salmonella spp., and 71.43 % and 100 % for Shiga toxins (stx), respectively. Overall sensitivity of the LAMP assay was 89.81 % for samples with PCR Ct values ≤40, and 95.14 % when using a Ct cut-off ≤35, respectively. More samples tested positive for C. jejuni, C. coli and stx1 by LAMP than by culture. There was 100 % concordance between the two methods for stx2 and Y. enterocolitica. Four out of 25 Salmonella cases were identified by culture but not by LAMP. With a test run time of 30 min and a few minutes for sample preparation, the LAMP assay could be useful for diagnosing bacterial gastrointestinal pathogens in individual cases where specific therapeutic decisions are required.
分子检测通常基于多重PCR技术,由于其结果更快且工作流程比传统培养更直接,越来越多地用于诊断细菌性肠胃炎。许多依赖培养的方法和半自动PCR检测劳动强度大,需要专业技术知识。因此,易于操作并能及时鉴定最常见肠道细菌病原体的检测方法可能会受到关注。本研究调查了一种基于环介导等温扩增(LAMP)的分子检测方法,用于快速鉴定几种常见细菌病原体。共分析了204份粪便样本。与作为参考方法的BD MAX™肠道细菌检测板PCR相比,该检测方法的灵敏度和特异性如下:弯曲杆菌属分别为88.35%和99.04%,沙门氏菌属分别为88%和100%,志贺毒素(stx)分别为71.43%和100%。对于PCR Ct值≤40的样本,LAMP检测的总体灵敏度分别为89.81%,使用Ct截止值≤35时为95.14%。通过LAMP检测出空肠弯曲菌、结肠弯曲菌和stx1呈阳性的样本比培养法更多。两种方法对stx2和小肠结肠炎耶尔森菌的检测结果100%一致。25例沙门氏菌病例中有4例通过培养法鉴定出来,但LAMP检测未鉴定出。LAMP检测的运行时间为30分钟,样本制备只需几分钟,在需要做出特定治疗决策的个别病例中,该检测方法可能有助于诊断细菌性胃肠道病原体。