Deletion of Salmonella enterica serovar typhimurium sipC gene

Objective:To construct a novel plasmid as Salmonella enterica serovar typhimurium(S.typhimurium)sip C gene knockouts candidate.Methods:In this research,50upstream and 30downstream regions of S.typhimurium sip C gene and kanamycin gene were PCR amplified.Each of these DNA fragment was cloned into p GEM T-easy vector.The construct was confirmed by PCR and restriction digest.Results:PCR amplified 320,206 and 835 bp DNA fragments were subcloned into p ET-32 vector resulting with a plasmid called p ET-32-sip C up-kan-sip C down.Conclusions:The new plasmid(p ET-32-sip C up-kan-sip C down)is useful for genetic engineering and for future manipulation of S.typhimurium sip C gene.

The relationship between <i>Helicobacter pylori</i>disease and bacterial count in stomach

Several studies showed significant relationships between bacterial counts and the severity and type of disease in patients. The aim of this study was to evaluate the relationship between Helicobacter pylori disease and bacterial count. In this study, 287 patients with dyspeptic symptoms were evaluated. Three variables including patient-reported data, clinical signs and bacterial load of gastric specimens were analyzed. Biopsy samples were collected from patients who were referred for endoscopies because of dyspeptic symptoms. Initially, the clinical status was evaluated and recorded by a questionnaire. DNA extraction was performed and H. pylori was analyzed for bacterial count by Real-time PCR assay and specific primers and probe. The variety range of bacteria in specimens was 104 to 1012. The results revealed that a greater relationships existed between 1012 and gastric cancer (p = 0.036), also 104 and acid reflux (p = 0.006) and vomiting (p = 0.047). Real-time PCR assay provides a highly accurate, rapid and precise method for detection of H. pylori and determination of progressive disease due to this bacterium.

The potential utility of nested PCR for investigation of Coxiella burnetii in Iranian snails

Objective:To detect the prevalence of Coxiella burnetii(C.burnetii)in two species of snails consisted of Lymnaea palustris(L.palustris)and Pomacea canaliculata(P.canaliculata)by using nested PCR method in Chaharmahel Va Bakhtiari Province which is located in the southwest of Iran.Methods:A total of 160 snail samples consisted of 100 L.palustris and 60 P.canaliculata were collected from 4 rice paddy fields in the southwest of Iran between June and August 2014.Snails'DNA was extracted by a genomic DNA purification kit according to the manufacturer's instructions.Detection of the presence of C.burnetii's DNA was carried out by using a nested PCR assay with[specific primers outer membrane protein 1(OMP1)-OMP2 and OMP3-OMP4]targeting the com1 gene.Results:In this study,a total of 160 snail samples were tested and 15(9.37%)samples were found positive for C.burnetii,15 samples were positive from the L.palustris and there were no positive samples from P.canaliculata.Conclusions:Snails are kind of gastropods which seem to be harmless in life,but these small gastropods can be very dangerous for farmers,especially in humid climates.Also,C.burnetii in snails showed that this bacterium can be a factor of transmission of contamination to human beings and animals.