This study assessed the application of metagenomic next-generation sequencing in pathogen detection of periprosthetic joint infections. A total of 95 cases who previously had undergone hip and knee replacement undergoing revision from January 2018 to January 2021 were included in this study. Specimens of synovial fluid and deep-tissue were collected for culture and metagenomic next-generation sequencing, and patients were retrospectively categorized as infected or aseptic using the Musculoskeletal Infection Society criteria after revision surgery. The sensitivity, specificity, positive and negative predictive values were compared. A total of 36 cases had positive culture results and 59 cases had positive metagenomic next-generation sequencing results. Culture was positive in 34 infected cases (58.6%) and 2 aseptic cases (5.4%). Metagenomic next-generation sequencing was positive in 55 infected cases (94.8%) and 4 aseptic cases (10.8%). Five cases diagnosed with infection had other potential pathogens detected by metagenomic next-generation sequencing. Among the 24 culture-negative periprosthetic joint infections, metagenomic next-generation sequencing was able to identify potential pathogens in 21 cases (87.5%). From sampling to reporting, the average time needed for culture was 5.2 (95% CI 3.1–7.3) days, while that for metagenomic next-generation sequencing was 1.3 (95% CI 0.9–1.7) days. Metagenomic next-generation sequencing is more advantageous in pathogen detection of periprosthetic joint infection after total joint replacement, especially in patients with multiple infections or negative culture results.

Gray mold disease, caused by Botrytis cinerea, severely impacts grape production worldwide. Although proanthocyanidins (PAs) contribute to fungal pathogen resistance, their role in grape defense against B. cinerea remains unclear. Here, we demonstrate that VvMYBPA1, a key transcriptional regulator of PA biosynthesis, negatively modulates B. cinerea resistance in grape berries. While infection suppressed endogenous VvMYBPA1, its agroinfiltration-mediated transient overexpression in berries elevated susceptibility, paralleling reduced β-1,3-glucanase (BGL) and polyphenol oxidase (PPO) activities. Additionally, VvMYBPA1 overexpression elevated VvRBOHs’ expression and reduced peroxidase (POD) activity, resulting in excessive hydrogen peroxide (H₂O₂) accumulation and more cell death. Our results reveal that VvMYBPA1 negatively regulates B. cinerea resistance by disrupting antioxidant enzyme activity and ROS homeostasis, providing new insights into the interplay between PA biosynthesis and fungal defense mechanisms.