Association of coagulase-negative staphylococci with orthopedic infections detected by in-house multiplex real-time PCR

Clinical significance of coagulase-negative staphylococci (CoNS) has been gradually acknowledged in both healthcare and clinical research, but approaches for their precise discrimination at the species level remain scarce. The current study aimed to evaluate the association of CoNS with orthopedic infections, where accurate and prompt identification of etiology is crucial for appropriate diagnosis and treatment decision-making. A 16S rRNA-based quantitative PCR (qPCR) assay was developed for the detection of genus and two panels of 3-plex qPCR assays for further differentiation of six CoNS species with remarkable clinical significance, including , , , , , and . All the assays exhibited excellent analytical performance. ΔCq (quantification cycle) between 16S rRNA and CoNS species-specific targets was established to determine the primary CoNS. These methods were applied to detect CoNS in wound samples from orthopedic patients with and without infection. Overall, CoNS were detected in 17.8% (21/118) of patients with clinically suspected infection and in 9.8% (12/123) of patients without any infection symptom (