First Experience of a Multicenter External Quality Assessment of Molecular 16S rRNA Gene Detection in Bone and Joint Infections

Abstract

The objective of this study was to assess the performance of seven French laboratories for 16S rRNA gene detection by real-time PCR in the diagnosis of bone and joint infection (BJI) to validate a large multicenter study. External quality control (QC) was required owing to the differences in extraction procedures and the molecular equipment used in the different laboratories. Three proficiency sets were organized, including four bacterial DNA extracts and four bead mill-pretreated osteoarticular specimens. Extraction volumes, 16S rRNA gene primers, and sequencing interpretation rules were standardized. In order to assess each laboratory's ability to achieve the best results, scores were assigned, and each QC series was classified as optimal, acceptable, or to be improved. A total of 168 QCs were sent, and 160 responses were analyzed. The expected results were obtained for 93.8%, with the same proportion for extracts (75/80) and clinical specimens (75/80). For the specimens, there was no significant difference between manual and automated extraction. This QC demonstrated the ability to achieve good and homogeneous results using the same 16S rRNA gene PCR with different equipment and validates the possibility of high-quality multicenter studies using molecular diagnosis for BJI.

The DendrisCHIP® Technology as a New, Rapid and Reliable Molecular Method for the Diagnosis of Osteoarticular Infections

“…The results obtained with our technology showed that in only 10% of the samples (34 out of 333) no bacteria were detected, and in the remaining 90%, 1 to 5 bacteria could be detected per sample, with a peak of 2 bacteria per sample in 40% of the total samples ( Figure 1 A). This result is in agreement with other molecular biology techniques that often detect more than one bacterium in bone and joint infections [ 13 , 42 , 43 ].…”

An update about molecular biology techniques to detect orthopaedic implant-related infections

“…Custom-made methodologies, based mainly in 16S rDNA amplification and sequencing, have high sensitivity and specificity, 38 , 39 and even high reproducibility is described in some reports. 40 , 41 These later studies are extremely important, because a well-known claimed limitation of this technology is the lack of reproducibility. In the study by Plouzeau et al 40 , 41 a control was submitted to different laboratories that have previously published the multicentre study.…”

“… 40 , 41 These later studies are extremely important, because a well-known claimed limitation of this technology is the lack of reproducibility. In the study by Plouzeau et al 40 , 41 a control was submitted to different laboratories that have previously published the multicentre study. 40 , 41 The results of reproducibility showed that, in well-trained experienced laboratories, this approach could be very useful and reproducible.…”

“…In the study by Plouzeau et al 40 , 41 a control was submitted to different laboratories that have previously published the multicentre study. 40 , 41 The results of reproducibility showed that, in well-trained experienced laboratories, this approach could be very useful and reproducible. Moreover, because this method could identify almost all existing bacteria, it has a very high potential for diagnosis.…”

Histopathological Diagnosis of Prosthetic Joint Infection: Does a Threshold of 23 Neutrophils Do Better than Classification of the Periprosthetic Membrane in a Prospective Multicenter Study?

“…Our West French network organization (Centre de Référence des Infections Ostéoarticulaires du Grand Ouest [CRIOGO]) for the multidisciplinary diagnosis and treatment of bone and joint infections in seven referral centers allowed us to carry out the first prospective multicenter study related to the molecular and histopathological diagnosis of PJI. The contribution of broad-range PCR to the diagnosis was recently published (17,18).…”