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通过 16S rDNA 扩增子的下一代测序鉴定感染性心内膜炎。

Identification of infective endocarditis by next-generation sequencing of 16S rDNA amplicons.

机构信息

Department of Laboratory Medicine, University of Washington Medical Center, Seattle, Washington 98195, USA.

Division of Infectious Diseases, Department of Medicine, California Pacific Medical Center, San Francisco, California 94115, USA.

出版信息

Cold Spring Harb Mol Case Stud. 2021 Feb 19;7(1). doi: 10.1101/mcs.a005876. Print 2021 Feb.

DOI:10.1101/mcs.a005876
PMID:33288524
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7903886/
Abstract

The oral aerotolerant anaerobe is an unusual cause of endocarditis and is amenable to treatment with β-lactam antibiotics. Because this organism is difficult to identify by conventional methods, molecular detection is a key diagnostic modality. Broad-range 16S rDNA PCR followed by Sanger sequencing constitute the first-line molecular approach, yet poor DNA quality, contaminating DNA, or low template quantity make identification challenging. Here we report a case of culture-negative, aortic and mitral valve endocarditis in a 66-yr-old woman with a history of cardiomyopathy, atrial fibrillation with intracardiac pacer, poor dentition, and recent tooth infection. In this case, 16S rDNA amplicon Sanger sequencing was not sufficient for pathogen identification because of interfering DNA, but deconvolution of the clinical sample using reflexive next-generation amplicon sequencing enabled confident identification of a single pathogenic organism, The patient developed a sigmoid colon perforation and died despite additional surgical treatment. Most endocarditis cases have been subacute and have been successfully treated with antibiotics, with or without valve replacement. This case highlights both an unusual etiologic agent of endocarditis, as well as the rational utilization of advanced molecular diagnostics tools for characterizing serious infections.

摘要

口腔耐气需氧菌是一种不常见的心内膜炎病因,对β-内酰胺类抗生素治疗有效。由于这种生物体很难通过常规方法识别,因此分子检测是一种关键的诊断方式。广泛的 16S rDNA PCR 随后进行 Sanger 测序构成了一线分子方法,但 DNA 质量差、污染 DNA 或模板数量低使得鉴定具有挑战性。我们在这里报告了一例 66 岁女性的培养阴性、主动脉瓣和二尖瓣心内膜炎病例,该患者有心肌病、心脏起搏器辅助的心房颤动、牙齿不良和近期牙齿感染史。在这种情况下,由于干扰 DNA,16S rDNA 扩增子 Sanger 测序不足以进行病原体鉴定,但通过使用反射性下一代扩增子测序对临床样本进行反卷积,能够确定单一的致病生物体。患者发生乙状结肠穿孔,尽管进行了额外的手术治疗,但仍死亡。大多数心内膜炎病例为亚急性,经过抗生素治疗(无论是否进行瓣膜置换术)都取得了成功。本病例既突出了心内膜炎的一种不常见病因,也强调了合理利用先进的分子诊断工具来描述严重感染。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5aa7/7903886/242e103d925d/MCS005876Lie_F2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5aa7/7903886/1aa032b5fd31/MCS005876Lie_F1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5aa7/7903886/242e103d925d/MCS005876Lie_F2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5aa7/7903886/1aa032b5fd31/MCS005876Lie_F1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5aa7/7903886/242e103d925d/MCS005876Lie_F2.jpg

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