URMITE CNRS-IRD UMR 6236, Faculté de Médecine et de Pharmacie, Université de la Méditerranée, 27 Bd Jean Moulin, 13385, Marseille Cedex 05, France.
Eur J Clin Microbiol Infect Dis. 2009 Nov;28(11):1363-8. doi: 10.1007/s10096-009-0793-6. Epub 2009 Aug 14.
The aim of this study was to compare 16 S rRNA gene amplification and sequencing with a systematic real-time PCR assay screening strategy that includes all common known pathogens recovered from lymph node biopsy specimens. Lymph node biopsy samples sent to our laboratory from January 2007 to December 2008 were tested in the study. Lymph nodes were screened for the presence of any bacteria by PCR amplification and sequencing targeting the 16 S rRNA gene and also by a specific real-time PCR strategy that includes Bartonella henselae, mycobacteria, Francisella tularensis, and Tropheryma whipplei. By testing 491 lymph nodes, we found that the sensitivity of our specific real-time PCR assay strategy was significantly higher than 16 S rRNA PCR amplification and sequencing for the detection of Bartonella henselae (142 vs 98; p < 10(-4)), Francisella tularensis (16 vs 10, p < 10(-4)), and mycobacteria (8 versus 3, p < 10(-4)). None of the samples was positive for Tropheryma whipplei. Our study demonstrates the usefulness and specificity of a systematic real-time PCR strategy for molecular analysis of lymph node biopsy specimens and the higher sensitivity compared with standard 16 S rRNA gene amplification and sequencing.
本研究旨在比较 16S rRNA 基因扩增和测序与一种系统的实时 PCR 检测策略，该策略包括从淋巴结活检标本中回收的所有常见已知病原体。研究中测试了 2007 年 1 月至 2008 年 12 月期间送到我们实验室的淋巴结活检样本。通过针对 16S rRNA 基因的 PCR 扩增和测序以及包括汉赛巴尔通体、分枝杆菌、土拉弗朗西斯菌和嗜吞噬细胞无形体的特定实时 PCR 策略，对淋巴结进行了任何细菌的筛查。通过测试 491 个淋巴结，我们发现我们的特定实时 PCR 检测策略的灵敏度明显高于 16S rRNA PCR 扩增和测序，用于检测汉赛巴尔通体（142 对 98；p < 10(-4))、土拉弗朗西斯菌（16 对 10，p < 10(-4)) 和分枝杆菌（8 对 3，p < 10(-4))。没有一个样本对嗜吞噬细胞无形体呈阳性。我们的研究表明，系统的实时 PCR 策略对于淋巴结活检标本的分子分析是有用且特异的，并且与标准的 16S rRNA 基因扩增和测序相比具有更高的灵敏度。
