Akademik Veri Yönetim Sistemi
MIKROBIYOLOJI BULTENI, cilt.58, sa.2, ss.125-134, 2024 (SCI-Expanded)
The World Health Organization has included the problem of antibiotic resistance among the top 10
important health problems in the world. Treatment of infectious diseases has become more difficult due
to the spread of antibiotic resistance between bacteria via transposable elements. Vancomycin-resistant
enterococci (VRE) are of critical medical and public health importance due to their association with serious
nosocomial infections and high risk of death. One of the most important features of VREs is that they
have multiple antibiotic resistance and treatment options are reduced. Therefore, new treatment methods
are needed. The vanA gene constitutes the building block of the vancomycin resistance mechanism
and causes high resistance to vancomycin. In this study, it was aimed to investigate the neutralization
of the vancomycin resistance mechanism by creating vanA antisense RNA (asRNA). The vanA positive
VRE50 strain in our culture collection which was isolated from the clinical sample, was used to amplify
the vanA gene by polymerase chain reaction (PCR). The amplified vanA amplicon was inserted inversely
into the pUC19 plasmid by means of the enzyme cutting sites in the primers used. The resulting plasmid
was combined with the pAT392 plasmid which can replicate in gram-positive bacteria and a fusion
plasmid was created. The fusion plasmid whose orientation was confirmed, was transferred to the wild
strain VRE50 by electroporation method. Minimum inhibitory concentration (MIC) values of transformed
VRE (tVRE50) and wild type VRE50 strains used as control were determined by the E-Test method. The
vancomycin MIC value of the wild type VRE50 strain was determined as 1024 μg/mL and that of the
tVRE50 strain was 32 μg/mL and it was determined that the vancomycin resistance of the tVRE50 strain
decreased with asRNA (antisense RNA). Antisense RNA technology is an important method for neutralizing
the expression of genes. This study showed that neutralization of the vancomycin resistance gene
may provide a lower MIC value in a vancomycin-resistant enterococcus strain and lead to increased susceptibility.
This new approach provides a new method for VRE treatment by neutralizing the vancomycin
resistance mechanism. The result obtained in this study needs to be supported by in vivo tests.