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住院患者粪便多重 PCR 诊断的经验:肠致病性大肠杆菌(EPEC)和培养阴性弯曲杆菌的临床意义。

Experiences from multiplex PCR diagnostics of faeces in hospitalised patients: clinical significance of Enteropathogenic Escherichia coli (EPEC) and culture negative campylobacter.

机构信息

Department of Infectious Diseases, Akershus University Hospital, PO Box 1000, 1478, Lørenskog, Nordbyhagen, Norway.

Institute of Clinical Medicine, University of Oslo, Oslo, Norway.

出版信息

BMC Infect Dis. 2019 Jul 17;19(1):630. doi: 10.1186/s12879-019-4271-1.

Abstract

BACKGROUND

In hospitalised patients with diarrhoea a positive campylobacter stool Polymerase Chain Reaction (PCR) test with negative culture results as well as Enteropathogenic Escherichia coli (EPEC) positive stool PCRs, challenges the clinician and may lead the unexperienced clinician astray. The aim of the study was to elucidate the clinical significance of positive Campylobacter and/or EPEC test results in hospitalised patients with diarrhoea.

METHODS

We conducted a retrospective case-case study. Case groups with 1) EPEC only and 2) EPEC in combination with any other pathogen in the PCR multiplex array, 3) PCR positive/culture negative Campylobacter, and 4) PCR positive/culture positive Campylobacter were compared. Medical records were reviewed and cases classified according to pre-specified clinical criteria as infectious gastroenteritis or non-infectious causes for diarrhoea. We analyzed the association between laboratory findings (the 4 subgroups) and the pre-specified clinical classification. We further sequenced culture negative campylobacter samples and tested EPEC for bundle forming pilus A (bfpA) gene, distinguishing typical from atypical EPEC.

RESULTS

A total of 291 patients were included, 169 were PCR positive for Campylobacter and 122 for EPEC. For both pathogens, co-infections were more common in culture negative/PCR positive samples than in culture positive samples. Clinical characteristics differed significantly in and between groups. Campylobacter culture positive patients had very high prevalence of characteristics of acute infectious gastroenteritis, whereas patients with PCR positive test results only often had an alternative explanation for their diarrhoea. Culture positives were almost exclusively C. jejuni/coli, whereas in culture negatives, constituting a third of the total PCR positives, C. concisus was the most frequent species. The vast majority of EPEC only positives had documented non-infectious factors that could explain diarrhoea. The EPEC co-infected group mimicked the culture positive campylobacter group, with most patients fulfilling the infectious gastroenteritis criteria.

CONCLUSIONS

In hospitalised patients, positive PCR results for campylobacter and EPEC should be interpreted in a clinical context after evaluation of non-infectious diarrhoea associated conditions, and cannot be used as a stand-alone diagnostic tool.

摘要

背景

在住院腹泻患者中,粪便环介导恒温扩增(LAMP)检测阳性而培养阴性的弯曲菌,以及肠致病性大肠埃希菌(EPEC)粪便 PCR 阳性,会给临床医生带来挑战,可能会使缺乏经验的临床医生误入歧途。本研究旨在阐明住院腹泻患者粪便环介导恒温扩增检测阳性的弯曲菌和/或 EPEC 的临床意义。

方法

我们进行了一项回顾性病例对照研究。病例组包括 1)仅 EPEC 阳性,2)EPEC 与 PCR 多重阵列中任何其他病原体共同阳性,3)PCR 阳性/培养阴性的弯曲菌,和 4)PCR 阳性/培养阳性的弯曲菌。回顾病历并根据预先指定的临床标准将病例分类为感染性胃肠炎或非感染性腹泻。我们分析了实验室结果(4 个亚组)与预先指定的临床分类之间的关系。我们进一步对培养阴性的弯曲菌样本进行测序,并检测 EPEC 的束状菌毛 A 基因(bfpA),以区分典型和非典型 EPEC。

结果

共纳入 291 例患者,169 例弯曲菌环介导恒温扩增检测阳性,122 例 EPEC 阳性。对于这两种病原体,培养阴性/PCR 阳性样本中的合并感染比培养阳性样本更常见。组间和组内的临床特征差异显著。弯曲菌培养阳性患者的急性感染性胃肠炎特征非常普遍,而 PCR 阳性患者的腹泻常存在其他替代解释。培养阳性者几乎全部为空肠弯曲菌/大肠埃希菌,而在培养阴性者中,占总 PCR 阳性者的三分之一,最常见的物种是 C. concisus。绝大多数仅 EPEC 阳性患者有可解释腹泻的明确非感染性因素。EPEC 合并感染组与弯曲菌培养阳性组相似,大多数患者符合感染性胃肠炎标准。

结论

在住院患者中,应在评估非感染性腹泻相关疾病后,结合临床情况解释弯曲菌和 EPEC 的 PCR 阳性结果,不能将其作为独立的诊断工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29d8/6637723/25ad9ed5a426/12879_2019_4271_Fig1_HTML.jpg

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