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Objective To express Tp0751 laminin-binding adhesion of Treponema pallidum (T. pallidum) ,and assess the immunocompetence. Methods The Tp0751 ORF without upstream non-cod-ing region was ligated into the expression vector pET-28a( + ), and expressed in E. coli R2566. Its immuno-gen was analyzed by Western blot and ELISA. Results A fusion protein with molecular weight about 26×103 was attained after expression and purification. Western blot proved that the recombinant protein can specifically react with T. palliclum IgG positive sera. Specific humoral response were elicited after introducing recombinant protein in Zealand rabbit and the specific antibody titer was above 1:10 2400 detected by indi-rect ELISA. Conclusion The expressed recombinant protein showed excellent immunoeompetence, and the results lay the foundation for the research on its function to T. paUidum infection.
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OBJECTIVE To investigate the bacterial distribution and antibiotic resistance of main pathogens from urinary tract infection (UTI) in our hospital for the guidance of rational use of antibiotics. METHODS The antibiotic resistance of clinical pathogen isolates from urinary tract infection by routine bacterium culture from Jan 2006 to Dec 2007 were studied retrospectively. The extended spectrum ?-lactamases (ESBLs) were detected out to the Gram-negative bacilli. RESULTS There were 722 strains of pathogens in the whole 2773 urinary samples with the isolating rate 26.0%. Most of the Urinary tract infections of patients were caused by Gram-negative bacilli (69.4%),then by Gram-positive cocci (16.3%) and fungi (14.3%). The most common pathogens in urinary tract infection were Escherichia coli (47.9%),Enterococcus faecais (7.9%),E. faecium (4.0%),Proteus mirabilis (3.9%),and Pseudomonas aeruginosa (3.5%). The Gram-negative bacilli were found to be sensitive to imipenem and meropenem,but highly resistant to the most other antibiotics,while P. mirabilis was susceptive to amikacin and cephalosporins. The Gram-positive cocci were more sensitive to nitrofuvantoin and vancomycin,but highly resistant to penicillin,oxacillin and SMT. CONCLUSIONS E. coli is still the primary urinary pathogen among patients,and highly resistant to a lot of antibiotics,We should carry out cultivation,isolation,and antimicrobial susceptibility testing as soon as possible to guide reasonable clinical drug therapy.
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Objective To study the drug resistance and production status of highly producing AmpC enzymes, extended-spectrum β-lactamases (ESBLs) and drug resistance in Acinetobacter bau-mannii. Methods Drug sensitivity test was performed with Kirby-Bauer method. ESBLs were detected with phenotype confirmatory test (double-disk synergy test), and highly producing AmpC enzymes were measured with phenotype screening method. Results 93 strains of Acinetobacter baumannii were mainly obtained from respiratory department and neurosurgery, and isolated from sputum/pharyngeal stab, raw surface/wound secretion and urine samples. 15 strains (16.1%) of highly producing AmpCenzyme-producing Acinetobacter baumannii were detected, so were 10 strains (10. 8%) of ESBLs-pro-ducing isolates. All enzymes-producing strains displayed multi-resistant, and their resistant rates to the third cephalosporin, penicillins and aztreonam were much higher than enzymes non-producing ones. None was detected resistant to imipenem. Conclusion Occurrence of highly producing AmpC en-zymes and extended-spectrum β-lactamases (ESBLs) is one of causes for Acinetobacter baumannii multi-resistance. It suggests that detection of ESBLs-producing isolates should be strengthened so as to prevent their outbreak and spread.
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OBJECTIVE To develop the strong and the specific multi-epitope antigen for the exploiting diagnosis of Treponema pallidum.METHODS The immuno-dominant epitopes of Tp0453 and Tp17 were amplified by PCR respectively,and subcloned into the expression vector pQE32 to generate multi-epitopes recombinant plasmid pQE32/Tp0453-17.The recombinant protein was expressed in Escherichia coli M15.The immunoresponse of recombinant fusion protein was analyzed by Western blot.RESULTS The multi-epitopes recombinant plasmid was successfully constructed,enzyme digestion analysis and sequencing showed that the inserted target genes were Tp0453 and Tp17 gene,compared with the gene reported by GenBank,it had 100% similarity;SDS-PAGE analysis showed the recombinant plasmid could be expressed in M15,its relatively molecular mass(Mr) of expressed product was about 52.0?103.The Western blot result showed the recombinant protein could be recognized by anti-T.pallidum positive serum.CONCLUSIONS The expressed multi-epitopes recombinant antigen showed excellent immunoresponse.The results lay the foundation for research on development of quick diagnostic kit applying to detection of T.pallidum infection.