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Molecular Identification of Bacteria from Aseptically Loose Implants

机译:无菌疏松种植体中细菌的分子鉴定

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摘要

Polymerase chain reaction (PCR) assays have been used to detect bacteria adherent to failed orthopaedic implants, but some PCR assays have had problems with probable false-positive results. We used a combination of a Staphylococcus species-specific PCR and a universal PCR followed by DNA sequencing to identify bacteria on implants retrieved from 52 patients (92 implants) at revision arthroplasty. We addressed two questions in this study: (1) Is this method able to show the existence of bacterial DNA on presumed aseptic loosed implants?; and (2) What proportion of presumed aseptic or culture-negative implants was positive for bacterial DNA by PCR? Fourteen implants (15%) were believed infected, whereas 74 implants (85%) were believed aseptic. Each implant was sonicated and the resulting solution was submitted for dual real-time PCR assay and culture. All implants believed aseptically loose were culture-negative, but nine of the 74 (12%) had bacterial DNA by PCR; two (2.7%) were PCR-positive and also showed histologic findings suggestive of infection. Uniquely developed PCR and bacterial sequencing assays showed bacterial DNA on 12% of implants removed for presumed aseptic loosening. Additional studies are needed to determine the clinical importance of bacterial DNA detected by PCR but not by conventional culture.>Level of Evidence: Level III, diagnostic study. See the Guidelines for Authors for a complete description of levels of evidence.
机译:聚合酶链反应(PCR)测定法已被用来检测附着在失败的整形外科植入物上的细菌,但某些PCR测定法可能存在假阳性结果的问题。我们使用了葡萄球菌物种特异性PCR和通用PCR的组合,然后进行DNA测序,以鉴定翻修置换术中从52例患者(92例植入物)中获取的植入物上的细菌。我们在这项研究中解决了两个问题:(1)这种方法是否能够显示假定的无菌疏松植入物上细菌DNA的存在? (2)假定的无菌或培养阴性植入物中有多少比例的PCR细菌DNA呈阳性?据信十四个植入物(15%)被感染,而74个植入物(85%)被认为是无菌的。对每个植入物进行超声处理,然后将所得溶液进行双重实时PCR分析和培养。所有被认为无菌松动的植入物均为培养阴性,但74个中的9个(占12%)通过PCR获得了细菌DNA。其中两个(2.7%)的PCR阳性,并且还显示出提示感染的组织学发现。独特开发的PCR和细菌测序测定法显示12%的植入物上的细菌DNA被移去以进行无菌性松动。需要进行其他研究以确定通过PCR而非常规培养检测到的细菌DNA的临床重要性。>证据水平: III级,诊断性研究。有关证据水平的完整说明,请参见《作者指南》。

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