{"title":"高灵敏、快速检测肺炎克雷伯菌中 khe、blaKPC-2 和 blaNDM-1 基因的多重重组酶辅助 qPCR 分析法","authors":"Shao-wei Hua, Jie Wang, Zi-jin Zhao, Feng-yu Tian, Meng Zhao, Yu-xin Wang, Rui-qing Zhang, Zhi-qiang Han, Shi-jue Gao, Xiao-na Lv, Hong-yi Li, Xin-xin Shen, Xue-jun Ma, Zhi-shan Feng","doi":"10.1002/jcla.25038","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <h3> Objective</h3>\n \n <p>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<sub>KPC-2</sub>, and bla<sub>NDM-1</sub> genes in <i>Klebsiella pneumoniae</i>.</p>\n </section>\n \n <section>\n \n <h3> Methods</h3>\n \n <p>mRAP was carried out in a qPCR instrument within 1 h. The analytical sensitivities of mRAP for khe, bla<sub>KPC-2</sub>, and bla<sub>NDM-1</sub> 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.</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>mRAP achieved the sensitivities of 10, 8, and 14 copies/reaction for khe, bla<sub>KPC-2</sub>, and bla<sub>NDM-1</sub> genes, respectively, superior to qPCR. The Kappa value of qPCR and mRAP for detecting khe, bla<sub>KPC-2</sub>, and bla<sub>NDM-1</sub> genes was 1, 0.855, and 1, respectively (<i>p</i> < 0.05).</p>\n </section>\n \n <section>\n \n <h3> Conclusion</h3>\n \n <p>mRAP is a rapid and highly sensitive assay for potential clinical identification of khe, bla<sub>KPC-2</sub>, and bla<sub>NDM-1</sub> genes in <i>K. pneumoniae</i>.</p>\n </section>\n </div>","PeriodicalId":15509,"journal":{"name":"Journal of Clinical Laboratory Analysis","volume":"38 9","pages":""},"PeriodicalIF":2.6000,"publicationDate":"2024-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jcla.25038","citationCount":"0","resultStr":"{\"title\":\"A Multiplex Recombinase-Aided qPCR Assay for Highly Sensitive and Rapid Detection of khe, blaKPC-2, and blaNDM-1 Genes in Klebsiella pneumoniae\",\"authors\":\"Shao-wei Hua, Jie Wang, Zi-jin Zhao, Feng-yu Tian, Meng Zhao, Yu-xin Wang, Rui-qing Zhang, Zhi-qiang Han, Shi-jue Gao, Xiao-na Lv, Hong-yi Li, Xin-xin Shen, Xue-jun Ma, Zhi-shan Feng\",\"doi\":\"10.1002/jcla.25038\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n \\n <section>\\n \\n <h3> Objective</h3>\\n \\n <p>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<sub>KPC-2</sub>, and bla<sub>NDM-1</sub> genes in <i>Klebsiella pneumoniae</i>.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Methods</h3>\\n \\n <p>mRAP was carried out in a qPCR instrument within 1 h. The analytical sensitivities of mRAP for khe, bla<sub>KPC-2</sub>, and bla<sub>NDM-1</sub> 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.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Results</h3>\\n \\n <p>mRAP achieved the sensitivities of 10, 8, and 14 copies/reaction for khe, bla<sub>KPC-2</sub>, and bla<sub>NDM-1</sub> genes, respectively, superior to qPCR. The Kappa value of qPCR and mRAP for detecting khe, bla<sub>KPC-2</sub>, and bla<sub>NDM-1</sub> genes was 1, 0.855, and 1, respectively (<i>p</i> < 0.05).</p>\\n </section>\\n \\n <section>\\n \\n <h3> Conclusion</h3>\\n \\n <p>mRAP is a rapid and highly sensitive assay for potential clinical identification of khe, bla<sub>KPC-2</sub>, and bla<sub>NDM-1</sub> genes in <i>K. pneumoniae</i>.</p>\\n </section>\\n </div>\",\"PeriodicalId\":15509,\"journal\":{\"name\":\"Journal of Clinical Laboratory Analysis\",\"volume\":\"38 9\",\"pages\":\"\"},\"PeriodicalIF\":2.6000,\"publicationDate\":\"2024-04-08\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jcla.25038\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Clinical Laboratory Analysis\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/jcla.25038\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"MEDICAL LABORATORY TECHNOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Clinical Laboratory Analysis","FirstCategoryId":"3","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/jcla.25038","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MEDICAL LABORATORY TECHNOLOGY","Score":null,"Total":0}
A Multiplex Recombinase-Aided qPCR Assay for Highly Sensitive and Rapid Detection of khe, blaKPC-2, and blaNDM-1 Genes in Klebsiella pneumoniae
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, blaKPC-2, and blaNDM-1 genes in Klebsiella pneumoniae.
Methods
mRAP was carried out in a qPCR instrument within 1 h. The analytical sensitivities of mRAP for khe, blaKPC-2, and blaNDM-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, blaKPC-2, and blaNDM-1 genes, respectively, superior to qPCR. The Kappa value of qPCR and mRAP for detecting khe, blaKPC-2, and blaNDM-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, blaKPC-2, and blaNDM-1 genes in K. pneumoniae.
期刊介绍：
Journal of Clinical Laboratory Analysis publishes original articles on newly developing modes of technology and laboratory assays, with emphasis on their application in current and future clinical laboratory testing. This includes reports from the following fields: immunochemistry and toxicology, hematology and hematopathology, immunopathology, molecular diagnostics, microbiology, genetic testing, immunohematology, and clinical chemistry.
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