AUTHOR=Yu Lin , Kou Xianglan , Liu Ze , Guan Chushi , Sun Baoqing 

TITLE=Establishment and validation of a dual qPCR method for the detection of carbapenem-resistant Acinetobacter baumannii in bloodstream infections

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=Volume 15 - 2025

YEAR=2025

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2025.1490528

DOI=10.3389/fcimb.2025.1490528

ISSN=2235-2988

ABSTRACT=ObjectiveBloodstream infections(BSIs) caused by carbapenem-resistant Acinetobacter baumannii (CRAB) have a high mortality rate due to the high levels of drug resistance. There is an urgent need to establish a sensitive and accurate detection method to rapidly detect CRAB in BSIs.MethodsA new method was developed based on fluorescence quantitative PCR (qPCR) targeting the specific region of 16sRNA and OXA-23 gene from CRAB. The parameters were evaluated and optimized. This qPCR method was further applied in the detection of AB from 30 clinical specimens.ResultsThe qPCR method established in this study showed high specificity. The method successfully differentiated Acinetobacter baumannii(A. baumanii) from 26 other common pathogens in BSIs and identify the carbapenem resistance gene. The qPCR method shows a limit of detection (LOD) of 3×10-3 ng/μL, and displays good linear relationship between 16sRNA and OXA-23 and excellent repeatability (CV ≤2%). The results for the detection of 30 clinical specimens using this new qPCR method are in complete agreement with those using blood culture and drug susceptibility test.ConclusionThe qPCR method established in this study has strong specificity, wide linear range, good repeatability, and a lower LOD than PCR (Non-fluorescence quantification). The method provides new technical support for the early clinical diagnosis of CRAB in BSIs.