Palmer Michael P, Melton-Kreft Rachael, Nistico Laura, Hiller N Louisa, Kim Leon H J, Altman Gregory T, Altman Daniel T, Sotereanos Nicholas G, Hu Fen Z, De Meo Patrick J, Ehrlich Garth D
1 88th Surgical Operations Squadron, Orthopedic Surgery , Wright-Patterson Airforce Base, Dayton, Ohio.
2 Center of Excellence in Biofilm Research , Allegheny Health Network, Pittsburgh, Pennsylvania.
Genet Test Mol Biomarkers. 2016 Dec;20(12):721-731. doi: 10.1089/gtmb.2016.0080. Epub 2016 Oct 17.
Preliminary studies have identified known bacterial pathogens in the knees of patients with osteoarthritis (OA) before arthroplasty.
The current study was designed to determine the incidence and types of bacteria present in the synovial fluid of native knee joints from adult patients with diagnoses of septic arthritis and OA.
Patients were enrolled between October 2010 and January 2013. Synovial fluid samples from the affected knee were collected and evaluated with both traditional microbial culture and polymerase chain reaction-electrospray ionization-time-of-flight mass spectrometry (molecular diagnostics [MDx]) to prospectively characterize the microbial content. Patients were grouped by diagnosis into one of two cohorts, those with clinical suspicion of septic arthritis (n = 44) and those undergoing primary arthroplasty of the knee for OA (n = 21). In all cases where discrepant culture and MDx results were obtained, we performed species-specific 16S rRNA fluorescence in situ hybridization (FISH) as a confirmatory test.
MDx testing identified bacteria in 50% of the suspected septic arthritis cases and 29% of the arthroplasty cases, whereas culture detected bacteria in only 16% of the former and 0% of the latter group. The overall difference in detection rates for culture and MDx was very highly significant, p-value = 2.384 × 10. All of the culture-positive cases were typed as Staphylococcus aureus. Two of the septic arthritis cases were polymicrobial as was one of the OA cases by MDx. FISH testing of the specimens with discordant results supported the MDx findings in 91% (19/21) of the cases, including one case where culture detected S. aureus and MDx detected Streptococcus agalactiae.
MDx were more sensitive than culture, as confirmed by FISH. FISH only identifies bacteria that are embedded or infiltrated within the tissue and is thus not susceptible to contamination. Not all suspected cases of septic arthritis contain bacteria, but a significant percent of patients with OA, and no signs of infection, have FISH-confirmed bacterial biofilms present in the knee.
初步研究已在骨关节炎(OA)患者关节置换术前的膝关节中鉴定出已知的细菌病原体。
本研究旨在确定诊断为化脓性关节炎和OA的成年患者天然膝关节滑液中存在的细菌的发生率和类型。
患者于2010年10月至2013年1月入组。收集患侧膝关节的滑液样本,并采用传统微生物培养和聚合酶链反应 - 电喷雾电离 - 飞行时间质谱法(分子诊断[MDx])进行评估,以前瞻性地表征微生物成分。患者按诊断分为两个队列之一,即临床怀疑化脓性关节炎的患者(n = 44)和因OA接受膝关节初次置换术的患者(n = 21)。在所有获得不一致培养结果和MDx结果的病例中,我们进行了物种特异性16S rRNA荧光原位杂交(FISH)作为确证试验。
MDx检测在50%的疑似化脓性关节炎病例和29%的关节置换病例中检测到细菌,而培养仅在前一组的16%和后一组的0%中检测到细菌。培养和MDx检测率的总体差异非常显著,p值 = 2.384×10。所有培养阳性病例均被鉴定为金黄色葡萄球菌。MDx检测发现,2例化脓性关节炎病例为混合菌感染,1例OA病例也是如此。对结果不一致的标本进行FISH检测,在91%(19/21)的病例中支持MDx的结果,包括1例培养检测到金黄色葡萄球菌而MDx检测到无乳链球菌的病例。
如FISH所证实,MDx比培养更敏感。FISH仅识别嵌入或浸润在组织内的细菌,因此不易受到污染。并非所有疑似化脓性关节炎病例都含有细菌,但相当一部分无感染迹象的OA患者膝关节中存在FISH证实的细菌生物膜。