Duplex Reverse Transcription Multienzyme Isothermal Rapid Amplification Assays for Detecting SARS-CoV-2

Shang, Mei-Yun; Sun, Xian-Ge; Ning, Shunzong; Li, Bowen; Jiang, Yuansu; Chong, Huimin; Lu, Weiping; Qing, Huang

doi:10.7754/clin.lab.2021.210239

Cited by 4 publications

(1 citation statement)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The most representative technologies are RPA (recombinase polymerase amplification) and MIRA (multienzyme isothermal rapid amplification) ( Piepenburg et al., 2006 ; Lu et al., 2021 ). They can amplify complex DNA targets at room temperature within 10 min, which is more advantageous than other isothermal amplification methods ( Shang et al., 2021 ; Sun et al., 2021 ; Tu et al., 2022 ). In the detection format of MIRA amplifier, lateral flow dipstick (LFD) based MIRA detection has been widely established for various detection target ( Li et al., 2019a ; Li et al., 2021 ).…”

Section: Introductionmentioning

confidence: 99%

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

He¹,

Guo²,

Li³

2022

Front. Cell. Infect. Microbiol.

Self Cite

View full text Add to dashboard Cite

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.

show abstract