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Article Abstract
Objective: This study aimed to establish a highly sensitive and rapid single-tube, two-stage, multiplex recombinase-aided qPCR (mRAP) assay to specifically detect the khe, bla, and bla genes in Klebsiella pneumoniae.
Methods: mRAP was carried out in a qPCR instrument within 1 h. The analytical sensitivities of mRAP for khe, bla, and bla genes were tested using recombinant plasmids and dilutions of reference strains. A total of 137 clinical isolates and 86 sputum samples were used to validate the clinical performance of mRAP.
Results: mRAP achieved the sensitivities of 10, 8, and 14 copies/reaction for khe, bla, and bla genes, respectively, superior to qPCR. The Kappa value of qPCR and mRAP for detecting khe, bla, and bla genes was 1, 0.855, and 1, respectively (p < 0.05).
Conclusion: mRAP is a rapid and highly sensitive assay for potential clinical identification of khe, bla, and bla genes in K. pneumoniae.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11137842 | PMC |
| http://dx.doi.org/10.1002/jcla.25038 | DOI Listing |
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Article Synopsis
- * Our results showed that while no combination completely inhibited the rtRPA reaction, the autofluorescence depended significantly on the materials used and the specific wavelength tested.
- * Ultimately, we successfully detected specific genes using these 3D printed monolithic microreactors, achieving a high sensitivity comparable to traditional methods, and demonstrated that these devices could be manufactured for efficient rtRPA applications.