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应用 16S rRNA 基因 PCR 扩增技术检测发生十字韧带断裂的犬关节炎性膝关节中细菌 DNA 的季节性变化。

Seasonal variation in detection of bacterial DNA in arthritic stifle joints of dogs with cranial cruciate ligament rupture using PCR amplification of the 16S rRNA gene.

机构信息

Comparative Orthopaedic Research Laboratory, School of Veterinary Medicine, University of Wisconsin-Madison, 2015 Linden Drive, Madison, WI 53706, USA.

出版信息

Vet Microbiol. 2010 Feb 24;141(1-2):127-33. doi: 10.1016/j.vetmic.2009.08.015. Epub 2009 Aug 18.

Abstract

An underappreciated cause and effect relationship between environmental bacteria and arthritis may exist. Previously, we found that stifle arthritis in dogs was associated with the presence of environmental bacteria within synovium. Cranial cruciate ligament rupture (CCLR) is often associated with stifle arthritis in dogs. We now wished to determine whether seasonal variation in detection of bacterial material may exist in affected dogs, and to also conduct analyses of both synovium and synovial fluid. We also wished to analyze a larger clone library of the 16S rRNA gene to further understanding of the microbial population in the canine stifle. Synovial biopsies were obtained from 117 affected dogs from January to December 2006. Using PCR, synovium and synovial fluid were tested for Borrelia burgdorferi and Stenotrophomonas maltophilia DNA. Broad-ranging 16S rRNA primers were also used and PCR products were cloned and sequenced for bacterial identification. Overall, 41% of arthritic canine stifle joints contained bacterial DNA. Detection of bacterial DNA in synovial fluid samples was increased, when compared with synovium (p<0.01). Detection rates were highest in the winter and spring and lowest in the summer period, suggesting environmental factors influence the risk of translocation to the stifle. Organisms detected were predominately Gram's negative Proteobacteria, particularly the orders Rhizobiales (32.8% of clones) and Burkholderiales (20.0% of clones), usually as part of a polymicrobial population. PCR-positivity was inversely correlated with severity of arthritis assessed radiographically and with dog age. Bacterial translocation to the canine stifle may be associated with changes to the indoor environment.

摘要

关节内细菌与关节炎之间可能存在一种被低估的因果关系。此前,我们发现狗膝关节关节炎与滑膜内环境细菌的存在有关。十字韧带断裂(CCLR)常与狗膝关节关节炎有关。我们现在想确定受影响的狗是否存在细菌物质检测的季节性变化,同时对滑膜和滑液进行分析。我们还希望对更大的 16S rRNA 基因克隆文库进行分析,以进一步了解犬膝关节的微生物种群。2006 年 1 月至 12 月,我们从 117 只受影响的狗身上获得了滑膜活检。使用 PCR 检测滑膜和滑液中的伯氏疏螺旋体和嗜麦芽窄食单胞菌 DNA。还使用了广泛的 16S rRNA 引物,并对 PCR 产物进行克隆和测序以鉴定细菌。总的来说,41%的关节炎犬膝关节含有细菌 DNA。与滑膜相比,滑液样本中细菌 DNA 的检测率更高(p<0.01)。冬季和春季的检测率最高,夏季最低,表明环境因素影响到向膝关节转移的风险。检测到的生物主要是革兰氏阴性的变形菌门,特别是根瘤菌目(32.8%的克隆)和伯克霍尔德菌目(20.0%的克隆),通常是作为多微生物种群的一部分。PCR 阳性与影像学评估的关节炎严重程度和狗的年龄呈负相关。细菌向犬膝关节的转移可能与室内环境的变化有关。

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