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应用多重 PCR 检测方法对肾移植受者严重腹泻的微生物学诊断

Microbiological diagnosis of severe diarrhea in kidney transplant recipients by use of multiplex PCR assays.

机构信息

Clinical and Molecular Virology Unit, Faculty of Medicine, University Hospital and EA-4684 Cardiovir SFR-CAP sante, Reims, France.

出版信息

J Clin Microbiol. 2013 Jun;51(6):1841-9. doi: 10.1128/JCM.03366-12. Epub 2013 Apr 3.

Abstract

Diarrhea is a frequent complication after kidney transplantation, ascribed to adverse effects of the immunosuppressive therapy in case of negative microbiological examination of the stools. The aim of this study was to improve the microbiological diagnosis by implementing molecular tests. Fifty-four severe diarrhea events that occurred in 49 adult kidney transplant recipients from September 2010 to November 2011 were investigated. One or several enteric pathogens were detected in 13 (23%) stool samples using classical microbiological methods versus 39 (72%) for the seven commercially available multiplex PCR assays used retrospectively (P = 0.006). Interestingly, molecular diagnosis identified 15 multiple infections compared to none using classical techniques. The primary pathogens detected were enteropathogenic Escherichia coli (EPEC) (n = 15; 38%), Campylobacter spp. (n = 15; 38%), and Norovirus (n = 14; 36%). Specificities for Campylobacter and Norovirus infection diagnosis were 75 and 100%, respectively, by comparison to reference methods. Based on molecular findings, a cyclosporine-mycophenolate mofetil combination was identified as a risk factor for developing Norovirus-induced diarrhea. Norovirus infections were also responsible for higher weight loss than all the other causes of diarrhea. In samples from asymptomatic immunocompromised and immunocompetent patients, EPEC but not Norovirus and Campylobacter infections were detected at a frequency similar to that observed in symptomatic kidney transplant recipients. In conclusion, molecular tools significantly improved the detection of single and multiple enteric infections by comparison to classical techniques and could quickly become the key element in the management of severe acute diarrhea in transplant recipients.

摘要

腹泻是肾移植后的常见并发症,归因于免疫抑制治疗的不良反应,而粪便的微生物检查为阴性。本研究旨在通过实施分子检测来改善微生物学诊断。对 2010 年 9 月至 2011 年 11 月期间 49 例成年肾移植受者的 54 例严重腹泻事件进行了调查。与传统微生物学方法相比,7 种商业上可用于回顾性检测的多重 PCR 检测方法检测到 13 份(23%)粪便样本中存在 1 种或多种肠道病原体(P = 0.006),而用传统方法仅能检测到 39 份(72%)。有趣的是,分子诊断比传统技术多发现 15 例多重感染。主要病原体为肠致病性大肠杆菌(EPEC)(n = 15;38%)、弯曲菌属(n = 15;38%)和诺如病毒(n = 14;36%)。与参考方法相比,针对弯曲菌和诺如病毒感染诊断的特异性分别为 75%和 100%。基于分子发现,环孢菌素-霉酚酸酯联合用药被确定为诺如病毒诱导腹泻的危险因素。与所有其他腹泻病因相比,诺如病毒感染导致的体重减轻更为严重。在无症状免疫抑制和免疫功能正常的患者样本中,与在有症状的肾移植受者中观察到的情况一样,EPEC 而不是诺如病毒和弯曲菌感染的频率相似。总之,与传统技术相比,分子工具显著提高了对单一和多种肠道感染的检测,并且可能很快成为移植受者严重急性腹泻管理的关键因素。

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