Rapid Detection of New Delhi Metallo-β-Lactamase Gene Using Recombinase-Aided Amplification Directly on Clinical Samples From Children

Frontiers in Microbiology

Lin Gan¹  Guanhua Xue¹  Jinghua Cui¹  Weiwei Zhang¹  Nannan Li¹  Chao Yan¹  Jing Yuan¹  Rui Zhang¹  Junxia Feng¹  Shiyu Liu¹  Ting Zhang¹  Hanqin Zhao¹  Hailan Yao¹  Yanling Feng¹  Shuheng Du¹  Jun Tai²  Lijuan Ma²  Dong Qu²  Wenjian Xu²  Yongxiang Wei² 

[1] Capital Institute of Pediatrics, Beijing, China;Children’s Hospital Affiliated to Capital Institute of Pediatrics, Beijing, China;

关键词: carbapenemase;    blaNDM;    recombinase-aided amplification;    pediatrics;    character;   

DOI  :  10.3389/fmicb.2021.691289

来源: DOAJ

【 摘 要 】

New Delhi metallo-β-lactamase, a metallo-β-lactamase carbapenemase type, mediates resistance to most β-lactam antibiotics including penicillins, cephalosporins, and carbapenems. Therefore, it is important to detect blaNDM genes in children’s clinical samples as quickly as possible and analyze their characteristics. Here, a recombinase-aided amplification (RAA) assay, which operates in a single one-step reaction tube at 39°C in 5−15 min, was established to target blaNDM genes in children’s clinical samples. The analytical sensitivity of the RAA assay was 20 copies, and the various bacterial types without blaNDM genes did not amplify. This method was used to detect blaNDM genes in 112 children’s stool samples, 10 of which were tested positive by both RAA and standard PCR. To further investigate the characteristics of carbapenem-resistant bacteria carrying blaNDM in children, 15 carbapenem-resistant bacteria (Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Citrobacter freundii, Klebsiella oxytoca, Acinetobacter junii, and Proteus mirabilis) were isolated from the 10 samples. Notably, more than one bacterial type was isolated from three samples. Most of these isolates were resistant to cephalosporins, cefoperazone-sulbactam, piperacillin-tazobactam, ticarcillin-clavulanic acid, aztreonam, co-trimoxazole, and carbapenems. blaNDM–1 and blaNDM–5 were the two main types in these samples. These data show that the RAA assay has potential to be a sensitive and rapid blaNDM gene screening test for clinical samples. The common existence of blaNDM and multi-drug resistance genes presents major challenges for pediatric treatment.

【 授权许可】

Unknown