Colitis caused by Entamoeba histolytica identified by real-time-PCR and fluorescence in situ hybridization from formalin-fixed, paraffin-embedded tissue.

Andreas Müller, Hagen Frickmann, Egbert Tannich, Sven Poppert, Ralf Matthias Hagen
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Abstract

Intestinal amoebiasis in a 35-year-old German patient with a 3 weeks travel history in Indonesia was initially misidentified as non-steroidal anti-inflammatory-drug associated colitis in colonoscopy and histopathological analysis. Furthermore, initial stool examination by microscopy and Entamoeba faecal antigen ELISA did not reveal any protozoan infection. When cessation of non-steroidal anti-inflammatory drug (NSAID) use and mesalazine treatment did not lead to clinical improvement, the patient presented to a specialist for tropical diseases. An intensive reinvestigation including a workup of formalin-fixed, paraffin-embedded colonic biopsies by molecular analysis with real-time PCR and fluorescence in situ hybridization (FISH) proofed the diagnosis of Entamoeba histolytica colitis. Molecular methods including real-time PCR and FISH for the diagnosis of amoebiasis from histopathological samples are rarely used for the diagnosis of E. histolytica infections. Bloody diarrhoea vanished after the onset of metronidazole treatment. In conclusion, the here-presented case demonstrates how modern molecular diagnostics may help to diagnose E. histolytica-associated colitis, even from difficult specimens like paraffin-embedded, formalin-fixed tissue.

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通过实时聚合酶链式反应和荧光原位杂交技术从福尔马林固定石蜡包埋组织中鉴定出由组织溶解恩塔米巴虫引起的结肠炎。
一名 35 岁的德国患者曾在印度尼西亚旅行 3 周,其肠道阿米巴病最初在结肠镜检查和组织病理学分析中被误认为是非类固醇抗炎药物相关性结肠炎。此外,通过显微镜和恩塔米巴粪便抗原酶联免疫吸附法进行的初步粪便检查并未发现任何原生动物感染。由于停止使用非甾体抗炎药(NSAID)和美沙拉嗪(mesalazine)治疗仍未改善临床症状,患者前往热带病专科就诊。通过对福尔马林固定、石蜡包埋的结肠活检样本进行实时 PCR 和荧光原位杂交(FISH)分子分析等强化再调查,证实了恩塔莫阿组织溶解虫结肠炎的诊断。通过组织病理学样本诊断阿米巴病的分子方法(包括实时 PCR 和 FISH)很少用于诊断组织溶解埃塔巴虫感染。甲硝唑治疗后,血性腹泻消失了。总之,本病例展示了现代分子诊断技术如何帮助诊断溶组织埃希氏菌相关性结肠炎,即使是从石蜡包埋、福尔马林固定的组织等困难标本中也能诊断出来。
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