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实时定量逆转录聚合酶链反应检测中国结节病患者淋巴结中丙酸杆菌核糖体RNA

Real-time quantitative reverse transcription-polymerase chain reaction to detect propionibacterial ribosomal RNA in the lymph nodes of Chinese patients with sarcoidosis.

作者信息

Zhou Y, Wei Y-R, Zhang Y, Du S-S, Baughman R P, Li H-P

机构信息

Department of Respiratory Medicine, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, China.

Department of Medicine, University of Cincinnati Medical Center, Cincinnati, OH, USA.

出版信息

Clin Exp Immunol. 2015 Sep;181(3):511-7. doi: 10.1111/cei.12650. Epub 2015 Jul 1.

Abstract

The aim of this study was to investigate the diagnostic value of using the copy number of propionibacterial rRNA as a biomarker for sarcoidosis. Ribosomal RNA of Propionibacterium acnes and P. granulosum was measured by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) using formalin-fixed and paraffin-embedded tissue of lymph node biopsy from 65 Chinese patients with sarcoidosis, 45 with tuberculosis and 50 controls with other diseases (23 with non-specific lymphadenitis and 27 with mediastinal lymph node metastasis from lung cancer). The receiver operating characteristic (ROC) curve was analysed to determine an optimal cut-off value for diagnosis, and the diagnostic accuracy of the cut-off value was evaluated in additional tissue samples [24 patients with sarcoidosis and 22 with tuberculosis (TB)]. P. acnes or P. granulosum rRNA was detected in 48 of the 65 sarcoidosis samples but only in four of the 45 TB samples and three of the 50 control samples. Analysis of the ROC curve revealed that an optimal cut-off value of the copy number of propionibacterial rRNA for diagnosis of sarcoidosis was 50·5 copies/ml with a sensitivity and specificity of 73·8 and 92·6%, respectively. Based on the cut-off value, 19 of the 24 additional sarcoidosis samples exhibited positive P. acnes or P. granulosum, whereas only one of the 22 additional TB samples was positive, resulting in a sensitivity and specificity of 79·2 and 95·5%, respectively. These findings suggest that propionibacteria might be associated with sarcoidosis granulomatous inflammation. Detection of propionibacterial rRNA by RT-PCR might possibly distinguish sarcoidosis from TB.

摘要

本研究旨在探讨将丙酸杆菌rRNA的拷贝数作为结节病生物标志物的诊断价值。采用实时定量逆转录聚合酶链反应(RT-PCR)检测65例中国结节病患者、45例结核病患者及50例其他疾病对照者(23例非特异性淋巴结炎患者和27例肺癌纵隔淋巴结转移患者)经福尔马林固定、石蜡包埋的淋巴结活检组织中痤疮丙酸杆菌和颗粒丙酸杆菌的核糖体RNA。分析受试者工作特征(ROC)曲线以确定诊断的最佳临界值,并在额外的组织样本[24例结节病患者和22例结核病患者]中评估该临界值的诊断准确性。65例结节病样本中有48例检测到痤疮丙酸杆菌或颗粒丙酸杆菌rRNA,而45例结核病样本中仅4例、50例对照样本中仅3例检测到。ROC曲线分析显示,丙酸杆菌rRNA拷贝数诊断结节病的最佳临界值为50.5拷贝/ml,敏感性和特异性分别为73.8%和92.6%。基于该临界值,24例额外的结节病样本中有19例痤疮丙酸杆菌或颗粒丙酸杆菌呈阳性,而22例额外的结核病样本中仅1例呈阳性,敏感性和特异性分别为79.2%和95.5%。这些发现表明丙酸杆菌可能与结节病肉芽肿性炎症相关。通过RT-PCR检测丙酸杆菌rRNA可能有助于区分结节病和结核病。

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