ABSTRACT
Toxoplasma gondii is an obligatory intracellular protozoan. Considering to high prevalence of this disease the best way to reduce the raised loses is prevention of human and animal infection, rapid diagnosis, differentiation between acute and chronic disease. Rhoptry protein 1 of Toxoplasma gondii is an excretory-secretory antigen that exists in the most stages of life cycle. According to specifications of excretory-secretory antigen that seems this antigen is a suitable candidate to produce recombinant vaccine and diagnostic kit. The main object of the present work was cloning rhoptry protein 1 [ROP1] gene of Toxoplasma gondii [RH] in a cloning vector for further production of rhoptry proteins. Genomic DNA was extracted by phenol-chloroform method. The ROP1 fragment was amplified by PCR. This product was approved by sequencing and was cloned between the EcoR1 and Sal1 sites of the pTZ57R/T vector. Then transformed into Escherichia coli DH5 alpha strain and screened by IPTG and X-Gal. After isolating of this gene from pTZ57R/T, it was subcloned into pET32a plasmid. The plasmid was purified and approved by electrophoresis, enzyme restriction and PCR. After isolating of this gene from pTZ57R/T, it was subcloned into pET32a plasmid. After enzyme restriction and electrophoresis a fragment about 1183bp was separated from pET32a. Recombinant plasmid of ROP1 gene was constructed and ready for future study. That seems the antigen is a suitable candidate to produce recombinant vaccine and diagnostic kit
ABSTRACT
Pseudomonas aeruginosa is one of important causes of nosocomial infections that is an important medical problem in developed and developing countries. Resistance of this gram-negative organism towards various antibiotics, particularly beta-lactam and carbapenem has been reported increasingly. This study was done aiming at determining antibiotic resistance pattern and the frequency of extended-spectrum beta-lactamase in clinically isolated P. aeruginosa strains. In this cross-sectional study, 108 P. aeruginosa strains were collected from medical centers of Arak city. Antibiotic susceptibility of the isolated strains to antibiotics, such as, ceftazidime, imipenem, meropenem, ciprofloxacin, amikacin, and gentamicin was determined using disk diffusion method [Kirby-Bauer]. Identification of ESBL producing strains was performed by combined disk method. Also, MIC test was done on 36 isolates with four antibiotics including imipenem, ceftazidime, cefipime, and ciprofloxacin. Among the 108 isolates of P. aeruginosa, resistances to ceftazidime [33.3%], imipenem [22.2%], meropenem [24%], amikacin [20.3%], ciprofloxacin [15.7%] and gentamicin [19.4%] were obtained. In the MIC test, the rates of resistance to antibiotic were respectively reported 15, 20, 10, and 15%. Thirty-two strains out of 36 ceftazidime resistant strains [88.8%], were detected ESBL-positive. The results of this study are indicative of a high level resistance against different antibiotics of among P. aeruginosa isolates. Therefore, it is necessary to use a more appropriate treatment protocols