ArticleMicroorganisms2022
Development and Evaluation of Duplex MIRA-qPCR Assay for Simultaneous Detection of Staphylococcus aureus and non-aureus Staphylococci
Article in Microorganisms, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 19 citations in OpenAlex.
- RPA Combined With CRISPR/Cas12a for Rapid and Ultrasensitive Detection Dual-Gene of Methicillin-Resistant Staphylococcus aureus (MRSA).Journal of molecular recognition : JMR · 2026Article
- Development of a MIRA-CRISPR/Cas12a-based nucleic acid detection system for the discrimination ofJournal of ginseng research · 2026Article
- Red and blue microspheres based immunoassay combined with RPA and Cas12a for dual-detection of nucleic acid from foodborne pathogens.Mikrochimica acta · 2026Article
- Advances in rapid detection technologies for zoonotic diseases: a one health-oriented review.Frontiers in cellular and infection microbiology · 2026Review
- Evaluation and Application of the MIRA-qPCR Method for Rapid Detection of Norovirus Genogroup II in Shellfish.Microorganisms · 2025Article
- Article
- A novel system with robust compatibility and stability for detectingMicrobiology spectrum · 2024Article
- Development and evaluation of a real-time multienzyme isothermal rapid amplification assay for rapid detection of Streptococcus pneumoniae.Scientific reports · 2024Article
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Authors and funding
9 authors at 4 institutions in 1 country.
Funding
Abstract
Staphylococcus spp., especially Staphylococcus aureus (S. aureus), is an important pathogen in hospital-acquired infection and food poisoning. Here, we developed a multienzyme isothermal rapid amplification combined with duplex quantitative PCR (duplex MIRA-qPCR) method, which can simultaneously detect the S. aureus species-specific conserved gene FMN-bgsfp and the Staphylococcus genus-specific conserved gene tuf. This assay enabled the amplification of DNA within 20 min at a constant temperature of 39 °C. Specificity analysis indicated that all nine common Staphylococcus species were positive and non-Staphylococcus spp. were negative for tuf gene, whereas S. aureus was positive, non-aureus Staphylococci species and non-Staphylococcus spp. were negative for FMN-bgsfp gene, suggesting that duplex MIRA-qPCR exhibited high specificity. Meanwhile, the sensitivity was tested and the limit of detection (LoD) was 3 × 102 CFU/mL. The coefficient variation values ranged from 0.13% to 2.09%, indicating that the assay had good repeatability. Furthermore, all the nine common Staphylococcus species (including S. aureus) could be detected from four kinds of simulated samples and the LoD of S. aureus was 8.56 × 103 CFU/mL. In conclusion, the duplex MIRA-qPCR has advantages of stronger specificity, lower detection threshold, shorter detection time, and simpler operation, which is an effective tool to detect S. aureus and non-aureus Staphylococci spp. infections rapidly.
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