Arciola Carla Renata, Campoccia Davide, Baldassarri Lucilla, Donati Maria Elena, Pirini Valter, Gamberini Simonetta, Montanaro Lucio
Research Unit on Implant Infections, Rizzoli Orthopedic Institute, Bologna, Italy.
J Biomed Mater Res A. 2006 Feb;76(2):425-30. doi: 10.1002/jbm.a.30552.
Biofilm-forming ability is increasingly being recognized as an important virulence factor in Staphylococcus epidermidis. This study compares three different techniques for the detection of biofilm-positive strains. The presence of icaA and icaD genes responsible for biofilm synthesis was investigated by a PCR method in a collection of 80 S. epidermidis strains isolated from orthopedic implant infections. The results from molecular analysis were compared with those obtained by two classic phenotypic methods, the Congo red agar (CRA) plate test and the microtiter plate test (MtP). Fifty-seven percent of all the examined strains were found icaA/icaD-positive, of which only three were not positive for CRA test. Differently, by the MtP method, 66% of the strains were found to be biofilm-producers but only a limited agreement with the PCR-method was noticeable because of the observation of (icaA/icaD+)/MtP- strains (8%) and of a surprising ambiguous result of (icaA/icaD-)/MtP+ strains (16%). The category of the weak biofilm-producers provided the highest contribution to these mismatching results (10%). The better agreement between the CRA plate test with the molecular detection of ica genes indicates the former as a reliable test for the phenotypic characterization of virulence of clinical isolates. However, MtP method remains a precious tool for the in vitro screening of different biomaterials for the adhesive properties using a reference strain.
生物膜形成能力日益被认为是表皮葡萄球菌的一种重要毒力因子。本研究比较了三种检测生物膜阳性菌株的不同技术。采用聚合酶链反应(PCR)方法,对从骨科植入物感染中分离出的80株表皮葡萄球菌进行检测,研究负责生物膜合成的icaA和icaD基因的存在情况。将分子分析结果与两种经典表型方法(刚果红琼脂平板试验和微量滴定板试验)所得结果进行比较。在所有检测菌株中,57%被发现icaA/icaD呈阳性,其中只有三株刚果红琼脂平板试验呈阴性。不同的是,通过微量滴定板试验方法,66%的菌株被发现是生物膜产生菌,但由于观察到(icaA/icaD+)/微量滴定板试验阴性菌株(8%)以及令人惊讶的(icaA/icaD-)/微量滴定板试验阳性菌株(16%)的模糊结果,与PCR方法的一致性有限。弱生物膜产生菌类别对这些不匹配结果的贡献最大(10%)。刚果红琼脂平板试验与ica基因分子检测之间更好的一致性表明,前者是临床分离株毒力表型特征的可靠检测方法。然而,微量滴定板试验方法仍然是使用参考菌株对不同生物材料的粘附特性进行体外筛选的宝贵工具。