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

IF 2.6 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY Journal of Clinical Laboratory Analysis Pub Date : 2024-04-08 DOI:10.1002/jcla.25038

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

{"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}

引用次数: 0

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.

Abstract Image

查看原文

微信好友朋友圈 QQ好友复制链接

本刊更多论文

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

本研究旨在建立一种高灵敏度、快速的单管、两阶段、多重重组酶辅助qPCR（mRAP）检测方法，以特异性检测肺炎克雷伯菌中的khe、blaKPC-2和blaNDM-1基因。结果mRAP对khe、blaKPC-2和blaNDM-1基因的灵敏度分别为10、8和14拷贝/反应，优于qPCR。在检测 khe、blaKPC-2 和 blaNDM-1 基因方面，qPCR 和 mRAP 的 Kappa 值分别为 1、0.855 和 1（p < 0.05）。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文去求助

来源期刊

Journal of Clinical Laboratory Analysis 医学-医学实验技术

CiteScore

5.60

自引率

7.40%

发文量

584

审稿时长

6-12 weeks

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.