Journal
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
Volume 12, Issue -, Pages -Publisher
FRONTIERS MEDIA SA
DOI: 10.3389/fcimb.2022.1010201
Keywords
multienzyme isothermal rapid amplification; lateral flow dipstick; Acinetobacter baumannii; bloodstream infection; spiked blood specimens
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Funding
- Chongqing Natural Science Foundation
- Hospital Nurturing Fund of the First Affiliated Hospital of Chongqing Medical University
- Graduate Scientific Research and Innovation Project of Chongqing
- [cstc2020jcyj-msxmX0207]
- [PYJJ2019-216]
- [CYB21190]
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This study established a multienzyme isothermal rapid amplification with a lateral flow dipstick (MIRA-LFD) assay for detection of A. baumannii in spiked blood specimens. The developed MIRA-LFD assay showed high specificity and sensitivity and outperformed the real-time PCR method. It is a time-saving and effective tool for rapid and reliable diagnosis of A. baumannii bloodstream infection in primary hospitals.
PurposeThis study aimed to establish the multienzyme isothermal rapid amplification with a lateral flow dipstick (MIRA-LFD) assay and evaluate its performance in detection of A. baumannii in spiked blood specimens. MethodsThe study was divided into two stages: a pilot study to establish the methodology and a clinical validation study to evaluate its performance. In the first step, we designed primers specific to detect A. baumannii, optimized the MIRA-LFD assay and analyzed its performance regarding limits of detection, reproducibility, specificity, and efficiency of detection using real-time PCR method. In the second step, we obtained 50 spiked blood isolates and detected these pathogens by MIRA-LFD assay. The MIRA-LFD time was 15 min from DNA sample amplification to complete pathogen detection. ResultsThe developed MIRA-LFD assay displayed a detection limit of 6 CFU/mL for detecting A. baumannii, which was significantly better than that of real-time PCR method, and no cross-reactivity was observed in other non-A. baumannii studied. The results obtained with 50 spiked blood isolates suggested that the developed MIRA-LFD assay had high specificity and sensitivity for identifying A. baumannii. ConclusionsThis study demonstrates that the established MIRA-LFD assay is time-saving, more effective and sensitive, which may become a powerful tool for rapid and reliable diagnosis of bloodstream infection caused by A. baumannii in primary hospitals.
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