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A Multiplex Recombinase-Aided qPCR Assay for Highly Sensitive and Rapid Detection of khe, blaKPC -2, and blaNDM -1 Genes in Klebsiella pneumoniae.
Hua, Shao-Wei; Wang, Jie; Zhao, Zi-Jin; Tian, Feng-Yu; Zhao, Meng; Wang, Yu-Xin; Zhang, Rui-Qing; Han, Zhi-Qiang; Gao, Shi-Jue; Lv, Xiao-Na; Li, Hong-Yi; Shen, Xin-Xin; Ma, Xue-Jun; Feng, Zhi-Shan.
Afiliação
  • Hua SW; Graduate School, Hebei North University, Zhangjiakou, China.
  • Wang J; Department of Clinical Laboratory, Hebei General Hospital, Shijiazhuang, China.
  • Zhao ZJ; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
  • Tian FY; Department of Clinical Laboratory, Hebei General Hospital, Shijiazhuang, China.
  • Zhao M; Graduate School, Hebei North University, Zhangjiakou, China.
  • Wang YX; Department of Clinical Laboratory, Hebei General Hospital, Shijiazhuang, China.
  • Zhang RQ; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
  • Han ZQ; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
  • Gao SJ; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
  • Lv XN; Graduate School, Hebei Medical University, Shijiazhuang, China.
  • Li HY; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
  • Shen XX; Graduate School, Hebei Medical University, Shijiazhuang, China.
  • Ma XJ; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
  • Feng ZS; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
J Clin Lab Anal ; 38(9): e25038, 2024 May.
Article em En | MEDLINE | ID: mdl-38590133
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, 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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Beta-Lactamases / Reação em Cadeia da Polimerase Multiplex / Klebsiella pneumoniae Limite: Humans Idioma: En Revista: J Clin Lab Anal Assunto da revista: TECNICAS E PROCEDIMENTOS DE LABORATORIO Ano de publicação: 2024 Tipo de documento: Article País de afiliação: China
Texto completo
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Beta-Lactamases / Reação em Cadeia da Polimerase Multiplex / Klebsiella pneumoniae Limite: Humans Idioma: En Revista: J Clin Lab Anal Assunto da revista: TECNICAS E PROCEDIMENTOS DE LABORATORIO Ano de publicação: 2024 Tipo de documento: Article País de afiliação: China