Wilbrink B, van der Heijden I M, Schouls L M, van Embden J D, Hazes J M, Breedveld F C, Tak P P
Leiden University Hospital, The Netherlands.
Arthritis Rheum. 1998 Mar;41(3):535-43. doi: 10.1002/1529-0131(199803)41:3<535::AID-ART20>3.0.CO;2-4.
Bacteria are considered to be important in the pathogenesis of several forms of arthritis. The goal of this study was to apply the 16S ribosomal RNA (rRNA)-polymerase chain reaction method for the detection of bacterial DNA in synovial fluid (SF) and synovial tissue (ST) from inflamed joints.
Samples from 5 patients with septic arthritis and from 7 with osteoarthritis or arthritis secondary to joint trauma were used as controls. Samples from 6 patients with spondylarthropathy (SpA) and from 20 with undifferentiated arthritis (UA) were analyzed for the presence of bacterial DNA using universal 16S rRNA primers. Automated sequencing and comparative data analysis were performed to identify the species.
In the positive control group, the bacterial species cultured from the synovium could be identified in all cases. No bacterial DNA was detected in the SF and ST from patients in the negative control group. In 4 of 6 patients with SpA and 7 of 20 with UA, the analysis of joint samples revealed the presence of bacterial DNA. Sequence analysis indicated the presence of multiple species, which was confirmed by sequencing of cloned products.
When the the above techniques were used with a stringent regimen, we were able to demonstrate that it is possible to collect and analyze joint samples without contaminating bacterial DNA. The accumulation of phagocytic cells that contain bacterial DNA of various species could play a role in the pathogenesis of both SpA and UA.
细菌被认为在多种关节炎的发病机制中起重要作用。本研究的目的是应用16S核糖体RNA（rRNA）聚合酶链反应方法检测来自炎症关节的滑液（SF）和滑膜组织（ST）中的细菌DNA。
将5例化脓性关节炎患者以及7例骨关节炎或关节创伤继发关节炎患者的样本用作对照。使用通用16S rRNA引物分析6例脊柱关节炎（SpA）患者和20例未分化关节炎（UA）患者的样本中细菌DNA的存在情况。进行自动测序和比较数据分析以鉴定菌种。
在阳性对照组中，所有病例均可鉴定出从滑膜培养的细菌种类。阴性对照组患者的SF和ST中未检测到细菌DNA。在6例SpA患者中的4例以及20例UA患者中的7例中，关节样本分析显示存在细菌DNA。序列分析表明存在多种菌种，这通过克隆产物测序得到证实。
当以上述严格方案使用上述技术时，我们能够证明在不污染细菌DNA的情况下收集和分析关节样本是可行的。含有各种菌种细菌DNA的吞噬细胞的积累可能在SpA和UA的发病机制中起作用。
