Development and clinical validation of a dual ddPCR assay for detecting carbapenem-resistant Acinetobacter baumannii in bloodstream infections

• Published in: Frontiers in microbiology 2024-03, Vol.15, p.1338395-1338395
• Main Authors: Kou, Xiaoxia, Zhu, Detu, Zhang, Yandong, Huang, Liyan, Liang, Jiawei, Wu, Ziman, Liu, Ze, Guan, Chushi, Yu, Lin
• Format: Article
• Language: English
• Subjects: Acinetobacter baumannii; antibiotic resistance; bloodstream infection; carbapenemase; ddPCR; Microbiology
• ISSN: 1664-302X; 1664-302X
• DOI: 10.3389/fmicb.2024.1338395

( , AB) represents a major species of Gram-negative bacteria involved in bloodstream infections (BSIs) and shows a high capability of developing antibiotic resistance. Especially, carbapenem-resistant (CRAB) becomes more and more prevalent in BSIs. Hence, a rapid and sensitive CRAB detection method is of urgent need to reduce the morbidity and mortality due to CRAB-associated BSIs. A dual droplet digital PCR (ddPCR) reaction system was designed for detecting the antibiotic resistance gene OXA-23 and AB-specific gene . Then, the specificity of the primers and probes, limit of detection (LOD), linear range, and accuracy of the assay were evaluated. Furthermore, the established assay approach was validated on 37 clinical isolates and compared with blood culture and drug sensitivity tests. The dual ddPCR method established in this study demonstrated strong primer and probe specificity, distinguishing CRAB among 21 common clinical pathogens. The method showed excellent precision (3 × 10  ng/μL, CV