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

IF 4.6 2区医学 Q2 IMMUNOLOGY

Frontiers in Cellular and Infection Microbiology Pub Date : 2025-02-26 eCollection Date: 2025-01-01 DOI:10.3389/fcimb.2025.1490528

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

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引用次数: 0

Abstract

Objective: Bloodstream 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.

Methods: A 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.

Results: The 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.

Conclusion: The 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.

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来源期刊

Frontiers in Cellular and Infection Microbiology IMMUNOLOGY-MICROBIOLOGY

CiteScore

7.90

自引率

7.00%

发文量

1817

审稿时长

14 weeks

期刊介绍： Frontiers in Cellular and Infection Microbiology is a leading specialty journal, publishing rigorously peer-reviewed research across all pathogenic microorganisms and their interaction with their hosts. Chief Editor Yousef Abu Kwaik, University of Louisville is supported by an outstanding Editorial Board of international experts. This multidisciplinary open-access journal is at the forefront of disseminating and communicating scientific knowledge and impactful discoveries to researchers, academics, clinicians and the public worldwide. Frontiers in Cellular and Infection Microbiology includes research on bacteria, fungi, parasites, viruses, endosymbionts, prions and all microbial pathogens as well as the microbiota and its effect on health and disease in various hosts. The research approaches include molecular microbiology, cellular microbiology, gene regulation, proteomics, signal transduction, pathogenic evolution, genomics, structural biology, and virulence factors as well as model hosts. Areas of research to counteract infectious agents by the host include the host innate and adaptive immune responses as well as metabolic restrictions to various pathogenic microorganisms, vaccine design and development against various pathogenic microorganisms, and the mechanisms of antibiotic resistance and its countermeasures.