Aims Microbiological culture is a key element in the diagnosis of periprosthetic joint infection (PJI). However, cultures of periprosthetic tissue do not have optimal sensitivity. One of the main reasons for this is that microorganisms are not released from the tissues, either due to biofilm formation or intracellular persistence. This study aimed to optimize tissue pretreatment methods in order to improve detection of microorganisms. Methods From December 2017 to September 2019, patients undergoing revision arthroplasty in a single centre due to PJI and aseptic failure (AF) were included, with demographic data and laboratory test results recorded prospectively. Periprosthetic tissue samples were collected intraoperatively and assigned to tissue-mechanical homogenization (T-MH), tissue-manual milling (T-MM), tissue-dithiothreitol (T-DTT) treatment, tissue-sonication (T-S), and tissue-direct culture (T-D). The yield of the microbial cultures was then analyzed. Results A total of 46 patients were enrolled, including 28 patients in the PJI group and 18 patients in the AF group. In the PJI group, 23 cases had positive culture results via T-MH, 22 cases via T-DTT, 20 cases via T-S, 15 cases via T-MM, and 13 cases via T-D. Three cases under ongoing antibiotic treatment remained culture-negative. Five tissue samples provided the optimal yield. Any ongoing antibiotic treatment had a relevant influence on culture sensitivity, except for T-DTT. Conclusion T-MH had the highest sensitivity. Combining T-MH with T-DTT, which requires no special equipment, may effectively improve bacterial detection in PJI. A total of five periprosthetic tissue biopsies should be sampled in revision arthroplasty for optimal detection of PJI. Cite this article: Bone Joint Res 2021;10(2):96–104.
Background This study aimed to evaluate the effects of different pretreatment methods on the microbial yield from infectious tissues. Methods Strains of Staphylococcus aureus (SA), Escherichia coli (EC) and Candida albicans (CA) were used to construct single-surface, full-surface, and internal infection models in sterile pork tissue. Manual milling (MM), mechanical homogenization (MH), sonificated (SF), dithiothreitol (DTT), and direct culture (DC) were used to pretreat these tissues, the microbial yield from different pretreatment methods were recorded and compared. Moreover, periprosthetic tissues collected intraoperatively from periprosthetic joint infection (PJI) patients were used as a verification. Results The study showed that the microbial yield from MH pretreatment was significantly higher than that of MM (P < 0.01) and SF pretreatment method (P < 0.01). Furthermore, in the internal infection model, the microbial yield from MH group was also significantly higher than that of SF (P < 0.01), DTT (P < 0.01), and DC group (P < 0.01). Moreover, the number of bacterial colonies obtained from periprosthetic tissues pretreated by MH was significantly higher than pretreated by other pretreatment methods (P = 0.004). Conclusions The effects of MH and DTT in microbial yield were significantly higher than that of DC, SF and MM, and these methods can be used to process multiple tissue samples at the same time, which might further improve the diagnostic sensitivity of infectious disease.
This study aimed to evaluate the effects of different pretreatment methods on the degree of microbial recovery in infected tissues. Standard strains of Staphylococcus aureus (SA), Escherichia coli (EC) and Candida albicans (CA) were used to construct single-surface, full-surface, and internal infection models in sterile pork tissue. Manual milling (MM), mechanical homogenization (MH), ultrasonic lysis (UL), dithiothreitol (DTT), and direct culture (DC) were used to pretreat infection tissues, the ability of the different pretreatment methods to achieve pathogen recovery in the different bacterial infection models was compared. At the same time, periprosthetic tissues collected from periprosthetic joint infection (PJI) were pretreated with the same methods. We showed that regardless of whether the single-surface or full-surface infection model was used in SA, EC, and CA infection, the microbial acquisition in the MH group was significantly higher than that in the MM (P <0.01) and UL groups (P <0.01). In the internal infection model, the microbial acquisition of the MH group was significantly higher than that of the MM (P <0.01), UL (P <0.01), DTT (P <0.01), and DC groups (P <0.01). In the PJI cases, the number of bacterial colonies obtained by MH was significantly higher than that obtained by other pretreatment methods (P = 0.004). The effects of MH and DTT in microbial recovery were significantly better than that of DC, UL and MM, and these methods can be used to process multiple tissue samples at the same time, which can further improve the efficiency of clinical microbial diagnosis.
Background: This study aimed to evaluate the effects of different pretreatment methods on the microbial yield in infected tissues. Methods: Strains of Staphylococcus aureus (SA), Escherichia coli (EC) and Candida albicans (CA) were used to construct single-surface, full-surface, and internal infection models in sterile pork tissue. Manual milling (MM), mechanical homogenization (MH), sonificated (SF), dithiothreitol (DTT), and direct culture (DC) were used to pretreat tissues, the microbial yield from different pretreatment methods were compared. At the same time, periprosthetic tissues collected intraoperatively from periprosthetic joint infection (PJI) patients were pretreated with the same methods. Results: The study showed that the microbial yield from MH pretreatment was significantly higher than that of MM (P <0.01) and SF pretreatment method (P <0.01). Furthermore, in the internal infection model, the microbial yield from MH group was also significantly higher than that of SF (P <0.01), DTT (P <0.01), and DC group (P <0.01). Moreover, the number of bacterial colonies obtained from periprosthetic tissues pretreated by MH was significantly higher than pretreated by other pretreatment methods (P = 0.004). Conclusions: The effects of MH and DTT in microbial yield were significantly higher than that of DC, SF and MM, and these methods can be used to process multiple tissue samples at the same time, which might further improve the sensitivity of infectious disease.
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