ABSTRACT
To evaluate the molecular mechanism of fluoroquinolones resistance in Mycoplasma hominis (MH) clinical strains isolated from urogenital specimens. 15 MH clinical isolates with different phenotypes of resistance to fluoroquinolones antibiotics were screened for mutations in the quinolone resistance-determining regions (QRDRs) of DNA gyrase (gyrA and gyrB) and topoisomerase IV (parC and parE) in comparison with the reference strain PG21, which is susceptible to fluoroquinolones antibiotics. 15 MH isolates with three kinds of quinolone resistance phenotypes were obtained. Thirteen out of these quinolone-resistant isolates were found to carry nucleotide substitutions in either gyrA or parC. There were no alterations in gyrB and no mutations were found in the isolates with a phenotype of resistance to Ofloxacin (OFX), intermediate resistant to Levofloxacin (LVX) and Sparfloxacin (SFX), and those susceptible to all three tested antibiotics. The molecular mechanism of fluoroquinolone resistance in clinical isolates of MH was reported in this study. The single amino acid mutation in ParC of MH may relate to the resistance to OFX and LVX and the high-level resistance to fluoroquinolones for MH is likely associated with mutations in both DNA gyrase and the ParC subunit of topoisomerase IV.
Subject(s)
Humans , Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial , Fluoroquinolones/pharmacology , Mutation, Missense , Mycoplasma Infections/microbiology , Mycoplasma hominis/drug effects , Reproductive Tract Infections/microbiology , DNA Gyrase/genetics , DNA Topoisomerase IV/genetics , Mycoplasma hominis/genetics , Mycoplasma hominis/isolation & purificationABSTRACT
<p><b>OBJECTIVE</b>To develop a quantitative real-time polymerase chain reaction (PCR) for detecting Rickettsia prowazekii.</p><p><b>METHODS</b>Primers and TaqMan-MGB probes designed based on ompB gene of R. prowazekii, were used to develop this method.</p><p><b>RESULTS</b>For the quantitative real-time PCR, the relationship between the values of threshold cycle (Ct) and the DNA copy number was linear (r = 0.999) and the sensitivity was about 100 times higher than that of the nested PCR for detecting the same DNA sample. The results of the genomic DNA samples of other rickettsial and bacterial agents detected by real-time PCR were all negative. DNAs extracted from blood samples of guinea pig infected with R. prowazekii were examined by real-time PCR and the positive results were obtained from some of these samples. However, the results of some samples in nested PCR assay were all negative.</p><p><b>CONCLUSION</b>These results suggested that the real-time PCR was highly specific and sensitive for detection of R. prowazekii that was useful for the detection of tiny DNA of R. prowazekii in blood samples from patients suspected of having epidemic typhus.</p>