AUTHOR=Hu Wei-Wei , He Jian-Wei , Guo Shu-Liang , Li Jin 

TITLE=Development and evaluation of a rapid and sensitive multienzyme isothermal rapid amplification with a lateral flow dipstick assay for detection of Acinetobacter baumannii in spiked blood specimens

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=12

YEAR=2022

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2022.1010201

DOI=10.3389/fcimb.2022.1010201

ISSN=2235-2988

ABSTRACT=<sec><title>Purpose</title><p>This study aimed to establish the multienzyme isothermal rapid amplification with a lateral flow dipstick (MIRA-LFD) assay and evaluate its performance in detection of <italic>A. baumannii</italic> in spiked blood specimens.</p></sec><sec><title>Methods</title><p>The 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 <italic>A. baumannii</italic>, 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.</p></sec><sec><title>Results</title><p>The developed MIRA-LFD assay displayed a detection limit of 6 CFU/mL for detecting <italic>A. baumannii</italic>, which was significantly better than that of real-time PCR method, and no cross-reactivity was observed in other non-<italic>A. baumannii</italic> studied. The results obtained with 50 spiked blood isolates suggested that the developed MIRA-LFD assay had high specificity and sensitivity for identifying <italic>A. baumannii</italic>.</p></sec><sec><title>Conclusions</title><p>This 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 <italic>A. baumannii</italic> in primary hospitals.</p></sec>