Detection of blaKPC and blaNDM genes by duplex PCR with lateral flow dipsticks from sterile body fluid samples. (27th November 2021)

• Record Type: Journal Article
• Authors: Sun, N.; Gu, Y.Y.; Wang, J.; Zhang, L.; Chen, F.F.; Zhang, J.X.; Wang, W.P.; Li, X.J.
• Is Part Of: Letters in applied microbiology. Volume 74:Number 2(2022)
• Page Start: 220
• Page End: 227
• Publication Date: 2021-11-27
• Subjects: blaKPC -- blaNDM -- carbapenem resistance -- duplex PCR -- lateral flow dipstick; Microbiology -- Periodicals; 660.62
• DOI: 10.1111/lam.13603
• Languages: English
• ISSNs: 0266-8254

Abstract

Abstract: Duplex polymerase chain reaction with lateral flow dipsticks (duplex PCR‐LFD) was developed for the simultaneous detection of beta‐lactamase Klebsiella pneumoniae carbapenemase ( bla KPC ) and beta‐lactamase New Dehli metallo‐beta‐lactamase ( bla NDM ) genes in body fluid samples. This method was validated using well‐characterized isolates. The assessment of the specificity of duplex PCR‐LFD showed that there was no cross‐reactivity with other targets. The detection limit of the duplex PCR‐LFD assay was 20 CFU per ml for bla KPC and bla NDM . Among 177 sterile body fluid samples tested by the duplex PCR‐LFD assay, 40 were bla KPC ‐positive and five were bla NDM ‐positive. The results obtained from 122 corresponding Gram‐negative bacteria which were isolated from these clinical samples and tested by duplex PCR‐LFD assay showed that there were 37 strains carrying bla KPC genes in 40 bla KPC ‐positive samples and three strains carrying bla NDM genes in five bla NDM ‐positive samples. Statistical analysis indicated that there was no significant difference between the direct detection of bla KPC and bla NDM genes in clinical sterile body fluid samples and their corresponding clinical isolates. Therefore, duplex PCR‐LFD can be effective for the simultaneous detection of bla KPC and bla NDM in clinical isolates and directly from clinical samples, which may be helpful for the administration of appropriate antimicrobial treatment. Abstract : Significance and Impact of the Study: The duplex polymerase chain reaction with lateral flow dipsticks (duplex PCR‐LFD) assay described in this study was developed for the simultaneous detection of two important carbapenemase genes: beta‐lactamase Klebsiella pneumoniae carbapenemase ( bla KPC ) and beta‐lactamase New Dehli metallo‐beta‐lactamase ( bla NDM ). This assay provides an effective and low‐cost method to detect of bla KPC and bla NDM genes from sterile body fluid samples and their corresponding clinical isolates. It is feasible to detect both bla KPC and bla NDM directly from clinical samples which can be verified by corresponding strains from these samples, and can significantly shorten the delay caused by bacterial isolation and culture. This can be helpful in administering appropriate antimicrobial treatments and as an effective tool for epidemiological investigation of these two carbapenemase genes.