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1. A Multiplex Recombinase‐Aided qPCR Assay for Highly Sensitive and Rapid Detection of khe, blaKPC‐2, and blaNDM‐1 Genes in Klebsiella pneumoniae

Author

Hua, Shao‐wei, primary, Wang, Jie, additional, Zhao, Zi‐jin, additional, Tian, Feng‐yu, additional, Zhao, Meng, additional, Wang, Yu‐xin, additional, Zhang, Rui‐qing, additional, Han, Zhi‐qiang, additional, Gao, Shi‐jue, additional, Lv, Xiao‐na, additional, Li, Hong‐yi, additional, Shen, Xin‐xin, additional, Ma, Xue‐jun, additional, and Feng, Zhi‐shan, additional

Published
2024
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4. A Multiplex Recombinase-Aided qPCR Assay for Highly Sensitive and Rapid Detection of khe, bla KPC -2 , and bla NDM -1 Genes in Klebsiella pneumoniae.

Author

Hua SW, Wang J, Zhao ZJ, Tian FY, Zhao M, Wang YX, Zhang RQ, Han ZQ, Gao SJ, Lv XN, Li HY, Shen XX, Ma XJ, and Feng ZS

Subjects
Humans, Klebsiella Infections microbiology, Klebsiella Infections diagnosis, Sensitivity and Specificity, Real-Time Polymerase Chain Reaction methods, Bacterial Proteins genetics, Recombinases genetics, Recombinases metabolism, Klebsiella pneumoniae genetics, Klebsiella pneumoniae isolation & purification, beta-Lactamases genetics, Multiplex Polymerase Chain Reaction methods
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 KPC-2 , and bla NDM-1 genes in Klebsiella pneumoniae., Methods: mRAP was carried out in a qPCR instrument within 1 h. The analytical sensitivities of mRAP for khe, bla KPC-2 , and bla NDM-1 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 KPC-2 , and bla NDM-1 genes, respectively, superior to qPCR. The Kappa value of qPCR and mRAP for detecting khe, bla KPC-2 , and bla NDM-1 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 KPC-2 , and bla NDM-1 genes in K. pneumoniae., (© 2024 The Authors. Journal of Clinical Laboratory Analysis published by Wiley Periodicals LLC.)

Published
2024
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