PURPOSE: Helicobacter pylori is a widely distributed gram-negative bacterium that infects the human stomach and duodenum. HpaA is a H. pylori-specific lipoprotein that has been shown to be an effective protective antigen against H. pylori infection. HpaA of H. pylori as a vaccine antigen is fully competent for stimulation of immune responses. The aim of this project is cloning, expression, and purification flagellar sheath adhesion of H. pylori in Escherichia coli host by fast protein liquid chromatography (FPLC) as a vaccination target. MATERIALS AND METHODS: The hpaA gene was inserted into pET28a (+) as cloning and expression vectors respectively. The recombinant plasmid (pET-hpaA) was subjected to sequencing other than polymerase chain reaction (PCR) and digestion analysis. Protein expression was induced by adding 1 mM isopropyl-beta-D-thiogalactoside to cultures of E. coli strain BL21 transformed with pET-hpaA. Protein expression assessed with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Protein purification of flagellar sheath adhesion was by FPLC. RESULTS: The restriction endonuclease digestion, PCR amplification analysis showed that the hpaA gene of 730 bp was amplified from H. pylori DNA and sequencing analysis of the pET-hpaA confirmed the cloning accuracy and in frame insertion of hpaA fragment. SDS-PAGE analysis showed the expression of an approximately 29,000 Da protein. CONCLUSION: Sequencing results along with SDS-PAGE analysis confirms the expression of recombinant hpaA in the heterologous E. coli BL21. Conclusion A prokaryotic expression system for hpaA gene was successfully constructed. These results indicate that production of a specific recombinant protein is an alternative and potentially more expeditious strategy for development of H. pylori vaccine.
Chromatography, Liquid ; Clone Cells* ; Cloning, Organism* ; Digestion ; DNA ; DNA Restriction Enzymes ; Duodenum ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli* ; Escherichia* ; Helicobacter pylori* ; Helicobacter* ; Humans ; Lipoproteins ; Plasmids ; Polymerase Chain Reaction ; Sodium Dodecyl Sulfate ; Stomach ; Vaccination*

BACKGROUND: This study aimed to determine the prevalence and factors related to needle stick injuries (NSIs) and to assess related safety measures among a sample of Iranian nurses. METHODS: In this cross-sectional study, a random sample of 168 registered active nurses was selected from different wards of one of the hospitals of Shiraz University of Medical Sciences (SUMS). Data were collected by an anonymous questionnaire and a checklist based observational method among the 168 registered active nurses. RESULTS: The prevalence of NSIs in the total of work experience and the last year was 76% and 54%, respectively. Hollow-bore needles were the most common devices involved in the injuries (85.5%). The majority of NSIs occurred in the morning shift (57.8%) and the most common activity leading to NSIs was recapping needles (41.4%). The rate of underreporting NSIs was 60.2% and the major reasons for not reporting the NSIs were heavy clinical schedule (46.7%) and perception of low risk of infection (37.7%). A statistically significant relationship was found between the occurrence of NSIs and sex, hours worked/week, and frequency of shifts/month. CONCLUSION: The study showed a high prevalence of NSIs among nurses. Supportive measures such as improving injection practices, modification of working schedule, planning training programs targeted at using personal protective equipment, and providing an adequate number of safety facilities such as puncture resistant disposal containers and engineered safe devices are essential for the effective prevention of NSI incidents among the studied nurses.
Anonyms and Pseudonyms ; Appointments and Schedules ; Checklist ; Cross-Sectional Studies ; Education ; Humans ; Iran* ; Needles* ; Needlestick Injuries* ; Prevalence ; Punctures