Abstract：
Sonicating explanted prosthetic implants to physically remove biofilms is a recognized method for improving the microbiological diagnosis of prosthetic joint infection (PJI); however, chemical and enzymatic treatments have been investigated as alternative biofilm removal methods. We compared the biofilm dislodging efficacy of sonication followed by the addition of enzyme cocktails with different activity spectra in the diagnosis of PJI with that of the sonication of fluid cultures alone. Consecutive patients who underwent prosthesis explantation due to infection at our institution were prospectively enrolled for 1 year. The diagnostic procedure included the collection of five intraoperative tissue cultures, sonication of the removed devices, and conventional culture of the sonication fluid. The resulting sonication fluid was also treated with an enzyme cocktail consisting of homemade dispersin B (0.04 µg/mL) and proteinase K (Sigma; 100 µg/mL) for 45 minutes at 37°C. The resulting sonication (S) and sonication with subsequent enzymatic treatment (SE) fluids were plated for aerobic and anaerobic culture broth for 7 days (aerobic) or 14 days (anaerobic). Identification was performed by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (Bruker). We included 107 patients from whom a prosthetic implant had been removed, among which PJI was diagnosed in 36 (34%). The sensitivity of S alone was significantly greater than that of SE alone (82% vs 71%; P < 0.05). Four patients with PJI were positive after sonication alone but negative after sonication plus enzymatic treatment. The four microorganisms missed after the addition of the enzyme cocktail were Staphylococcus aureus, two coagulase-negative Staphylococci, and Cutibacterium acnes. In conclusion, sonication alone was more sensitive than sonication followed by enzymatic treatment. The combination of these two methods had no synergistic effect; in contrast, the results suggest that the combination of both dislodgment methods affects the viability of gram-positive microorganisms.
OBJECTIVE: While the potential of sonication and enzymes as biofilm dispersal agents has been previously described, the originality of our work resides in the combination of both methods, which is hypothesized to enhance the ability to remove biofilm and, therefore, improve the microbiological diagnosis of PJI.
OBJECTIVE: While the potential of sonication and enzymes as biofilm dispersal agents has been previously described, the originality of our work resides in the combination of both methods, which is hypothesized to enhance the ability to remove biofilm and, therefore, improve the microbiological diagnosis of PJI.
摘要：
超声移植假体植入物以物理去除生物膜是一种公认的改善人工关节感染（PJI）微生物学诊断的方法；然而，已经研究了化学和酶处理作为替代的生物膜去除方法。我们比较了超声处理后添加具有不同活性谱的酶混合物在PJI诊断中的生物膜去除功效与仅对流体培养物进行超声处理的效果。连续在我们机构因感染而接受假体移植的患者前瞻性招募1年。诊断程序包括收集五种术中组织培养物，对移除的装置进行超声处理，和超声处理液的常规培养。在37°C下,用由自制分散剂B(0.04μg/mL)和蛋白酶K(Sigma;100μg/mL)组成的酶混合物处理所得的超声处理流体45分钟。将所得的超声处理(S)和超声处理与随后的酶处理(SE)流体铺板用于好氧和厌氧培养液7天(好氧)或14天(厌氧)。通过基质辅助激光解吸电离-飞行时间质谱(Bruker)进行鉴定。我们纳入了107名患者，这些患者的假体植入物已被移除，其中PJI被诊断为36（34％）。单独S的敏感性明显高于单独SE的敏感性（82％vs71％；P<0.05）。仅超声治疗后，四名PJI患者呈阳性，但超声治疗加酶治疗后呈阴性。添加酶混合物后错过的四种微生物是金黄色葡萄球菌，两种凝固酶阴性葡萄球菌，和粉刺杆菌.总之,单独的超声处理比超声处理后的酶处理更敏感。这两种方法的结合没有协同效应；相比之下，结果表明，两种移位方法的组合会影响革兰氏阳性微生物的生存能力。
目的：虽然先前已经描述了超声处理和酶作为生物膜分散剂的潜力，我们工作的独创性在于两种方法的结合，它被假设为增强去除生物膜的能力，因此,提高PJI的微生物学诊断。
目的：虽然先前已经描述了超声处理和酶作为生物膜分散剂的潜力，我们工作的独创性在于两种方法的结合，它被假设为增强去除生物膜的能力，因此,提高PJI的微生物学诊断。
