RESUMO
Background: Increased incidence of resistance to beta-lactams among members of the family Enterobacteriaceae has been reported worldwide. Extended spectrum beta-lactamase [ESBL] producing Gram-negative bacteria are becoming a major global concern and usually harbor plasmid-mediated enzymes of the TEM, SHV, OXA, PER, and CTX-M types. The aim of this study is to determine the prevalence of ESBL-producing Enterobacteriaceae in Mansoura hospitals and to molecularly characterize the ESBL-related bla genes, including blaTEM and blaCTX-M
Methodology: A total of 40 E. coli, 30 K. pneumoniae and 30 Proteus isolates were studied for antibiotic susceptibility pattern using different betalactam antibiotics and for the presence of ESBLs by combination of double-disc approximation test and inhibitor-potentiated disc-diffusion test. Subsequently, the hyper variable regions of beta-lactamase-encoding genes were amplified and sequenced using dye termination Sanger methodology to study the genetic variation among the clinical isolates
Results: All E. coli isolates were resistant to ampicillin, amoxicillin/clavulanate and cefadroxil. Regarding K. pneumoniae, all isolates were resistant to ampicillin, amoxicillin/clavulanate, cefadroxil, cefoxitin, cefuroxime and cefotaxime. Concerning Proteus species, all isolates were resistant to ampicillin, cefadroxil, cefotriaxone, cefuroxime and cefoperazone. In contrast 95% of E. coli isolates 80% of K. pneumoniae isolates and 90% of Proteus isolates were sensitive to imipenem. The detection of ESBLs by double-disc approximation test and inhibitor-potentiated disc-diffusion test was quiet different. Double disc approximation method lacks sensitivity. It showed false negative results in nearly 92% of the isolates that were concerned positive ESBLs producers by inhibitor-potentiated disc-diffusion test. PCR amplification and sequencing analysis revealed the presence of CTX-M and TEM type ESBLs in the tested isolates and could accurately characterize different types of blaTEM and blaCTX-M among the clinical isolates
Conclusion: Combined use of the conventional ESBLs screening methods and the molecular amplification of the ESBLs encoding genes followed by PCR based sequencing method provides a very valuable tool for identification and characterization of ESBLs producing E. coli, K. pneumoniae, and Proteus clinical isolates
RESUMO
Background: Gram negative bacteria are responsible for numerous infectious diseases. These diseases can occur in and harm any part of the body, the skin, eyes and the nervous, cardiovascular, respiratory, gastrointestinal and urogenital systems
Methods: In the present study, some phenotypic and molecular typing techniques were applied on 108 strains of E. coli, 88 strains of Ps. aeruginosa and 8 strains of Serratia isolated from different clinical lesions in Mansoura University Hospitals, Egypt
Results: The distribution of antibiotic resistance among the isolated strains showed high incidence of resistance and imipenem was the most active antibiotic. Using the active pyocin typing, 72 strains of Ps. Aeruginosa could be typed into 35 pyotypes. Using PCR technique it was found that 84% of the 50 tested isolates were found to have at least one of the tested ESBLs. Also TolC and AcrA genes were present in all tested E. coli except 4 strains and did not present in Ps. aeruginosa except 4 strains. Plasmid profiles of 23 tested E. coli appear to be diverse. Also the prevalence of plasmids in 22 tested Ps. aeruginosa strains was lower than in tested E. coli therefore 59.1% of tested Ps. aeruginosa strains harbored plasmids. Using Pyrosequencing technique, the sequenced region of gyrA, gyrB and ParC were able to differentiate between the tested strains and neighbor-joining tree was constructed to determine relatedness between the isolated strains. Moreover, Molecular cloning of the whole sequence of bla-TEM, bla-SHV and bla-CTX-M was carried out experimentally to study the expression of these genes and determine which genes of them responsible for the resistance
Conclusion: Molecular-based typing methods of are more advantageous compared with phenotypic typing methods in terms of better discrimination and reproducibility. Significant genetic variation was observed among different strains represented by the diversity of their plasmid profiles. All molecular genetics methods for distinguishing organism subtypes are based on differences in the DNA sequence
RESUMO
To evaluate the different markers of ovarian reserve [AMH-Inhibin B, FSH and antral follicle count [AFC] in insulin dependent diabetes mellitus [IDDM]. We studied 30 patients with IDDM as study groups [10cases >/=32 years group IV and 20 cases 32 years group III and 12 cases <32 years group I. Serum levels of FSH, LH, inhibin B and AMH were measured at [days 1-7] of menstrual cycle and AFC was done by trans vaginal ultrasound. AMH levels were lower in IDDM patients than in controls > 32 years [2.35 +/- 2.2 versus 7.79 +/- 1.73 p<0.000]. Also IDDM groups showed lower levels of inhibin B. While there is no difference in the levels of FSH. AFC is valuable for the diagnoses of premature ovarian failure in IDDM. Compared with FSH [AMH and inhibin B] are more valuable for the diagnosis of premature ovarian aging in IDDM patients which developed earlier decline in the ovarian follicle pool compared with the healthy women and also AMH is more valuable than AFC for detection of premature ovarian failure in IDDM