When the ability to identify Candida to species level of phenotypic and PCR-RFLP methods was assessed, a great difference was found between these two methods. It may be argued that Msp I and Bln I restriction enzyme fragments can be used in the identification of medically important Candida species. Further studies are needed to develop this kind of restriction profile to be used in the identification of candidal strains.
SUMMARY Leptospirosis is still one of the most important health problems in developing
countries located in humid tropical and subtropical regions. Human infections are
generally caused by exposure to water, soil or food contaminated with the urine of
infected wild and domestic animals such as rodents and dogs. The clinical course of
leptospirosis is variable and may be difficult to distinguish from many other
infectious diseases. The dark-field microscopy (DFM), serology and nucleic acid
amplification techniques are used to diagnose leptospirosis, however, a distinctive
standard reference method is still lacking. Therefore, in this study, we aimed to
determine the presence of Leptospira spp., to
differentiate the pathogenic L. interrogans and the non-pathogenic
L. biflexa, and also to determine the sensitivity and specificity
values of molecular methods as an alternative to conventional ones. A total of 133
serum samples, from 47 humans and 86 cattle were evaluated by two conventional tests:
the Microagglutination Test (MAT) and the DFM, as well as three molecular methods,
the 16S rRNA-PCR followed by Restriction Fragment Lenght Polymorphism (RFLP) of the
amplification products 16S rRNA-PCR-RFLP, LipL32-PCR and OmpL1-PCR. In this study,
for L. interrogans, the specificity and sensitivity rates of the 16S
rRNA-PCR and the LipL32-PCR were considered similar (100% versus 98.25% and 100%
versus 98.68%, respectively). The OmpL1-PCR was able to classify L.
interrogans into two intergroups, but this PCR was less sensitive
(87.01%) than the other two PCR methods. The 16S rRNA-PCR-RFLP could detect
L. biflexa DNA, but LipL32-PCR and OmpL1-PCR could not. The 16S
rRNA-PCR-RFLP provided an early and accurate diagnosis and was able to distinguish
pathogenic and non-pathogenic Leptospira species, hence it may be
used as an alternative method to the conventional gold standard techniques for the
rapid disgnosis of leptospirosis.
BackgroundThe resistance of aminoglycosides in strains that produce beta-lactamase can be developed through the multidrug resistant encoding genes carried by common plasmids. Recently, the association between 16S rRNA methyltransferase resistance and beta-lactamase enzymes carried by the same plasmids has drawn increased attention from researchers, particularly the association in aminoglycoside-resistant strains with a minimum inhibitory concentration (MIC) of ≥ 256 µg/mL.ObjectivesWe aimed to investigate the co-existence of 16S rRNA methyltransferase and beta-lactamase genes in multidrug resistant (MDR) Klebsiella pneumoniae strains isolated from clinical samples.MethodsWe determined the molecular mechanisms of aminoglycoside resistance and its relationship with resistance to carbapenem and beta-lactam group antibiotics in 40 extended-spectrum beta-lactamase (ESBL)-positive carbapenem- and aminoglycoside-resistant K. pneumoniae strains. Multidrug resistant K. pneumoniae was isolated from various clinical samples in the faculty of medicine of Cukurova University, Turkey. First, the resistance of aminoglycoside and beta-lactam antibiotics was phenotypically investigated using the Kirby-Bauer disk diffusion test, double disk synergy test, and modified Hodge test. The MIC values of aminoglycoside were determined using the agar dilution method. Polymerase chain reaction was performed to detect the carbapenemases, ESBL, and 16S rRNA methyltransferase genes. The results were confirmed by a sequence analysis.ResultsTwenty K. pneumoniae strains showed resistance to amikacin, and 40 were resistant to gentamicin. The MIC value was found to be > 512 µg/mL in five amikacin-resistant strains and > 128 µg/mL in 10 gentamicin-resistant isolates. The rmtC gene, a type of 16S rRNA methyltransferase, was amplified in four isolates (MIC amikacin: > 512 µg/mL, gentamicin: > 128 µg/mL). Of these four isolates, three had the blaNDM-1 gene and all contained at least one ESBL gene.ConclusionsThis study demonstrated the co-existence of rmtC and blaNDM-1 genes for the first time in Turkey. The spread of this resistant type should be monitored and limited through molecular surveillance.
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