Kolistin direncinin moleküler tespitinde yanlış tanı: kolistine duyarlı Acinetobacter baumannii izolatlarında yanlış mcr-1-PCR pozitifliği

Abstract

Purpose: The aim of this study was to investigate the presence of the mcr-1 gene, which is responsible for colistin resistance, in carbapenem-resistant Gram-negative bacteria that cause difficult-to-treat infections in a research hospital in Turkey. Materials and Methods: The mcr-1 gene was examined using PCR in 103 carbapenem-resistant isolates, including 75 Acinetobacter baumannii, 19 Pseudomonas aeruginosa, and 9 Klebsiella pneumoniae. DNA sequencing was performed to confirm the mcr-1 positivity. Other antimicrobial resistance genes were investigated in isolates that were found to be mcr-1-positive by PCR and colistin-resistant isolates. Results: Four (3.9% of the 103 carbapenem-resistant isolates and 5.3% of the 75 A. baumannii isolates) A. baumannii isolates, all susceptible to colistin, were found to be mcr-1-positive by PCR, whereas mcr-1 was not detected in four colistin-resistant isolates, one in A. baumannii and three in K. pneumoniae. DNA sequencing analysis determined that none of the amplification products was the targeted fragment, but they matched more than 70% with the chromosomal DNA fragments of A. baumannii strains. Therefore, these results were considered false-positive. Although these false-positive isolates were susceptible to colistin, they were extensively drug-resistant (XDR). Two of them were found to carry blaOXA23-like and blaTEM genes, another blaOXA23-like, blaTEM and blaOXA48-like genes, and the fourth one to have blaOXA23-like and blaCTXM genes. Conclusion: Although the specificity of the primers used to detect the mcr-1 gene by PCR was reported as 100% in most studies, we concluded that PCR tests are insufficient yet to use alone or with antibiotic susceptibility tests in rapid routine diagnosis. Confirming at least PCR-positive samples using DNA sequence analysis would be appropriate for a certain period.