In this study, 50 Vibrio cholerae O1 serotype Inaba isolates were collected during several cholera outbreaks throughout Iran during the summer of 2005. The results of antibiotic susceptibility testing showed that 86, 84, 84 and 82 % of the isolates were resistant to streptomycin, chloramphenicol, co-trimoxazole and tetracycline, respectively. The strains were genotyped using randomly amplified polymorphic DNA (RAPD), PFGE and ribotyping techniques. PCR showed that 100, 98 and 98 % carried the ctx, zot and ace genes, respectively. Biochemical fingerprinting of the isolates using the PhenePlate (PhP) system showed a low diversity index level (0.755), suggesting that the strains were highly homogeneous. Among the strains, 100 and 96 % showed an identical ribotype and PFGE patterns, respectively. The two isolates showing different PFGE patterns also exhibited discrete PhP types. RAPD was able to discriminate the isolates into six distinct groups, suggesting some genetic dissimilarity was present among the strains. These ribotyping, PFGE and PhP techniques revealed the clonal dissemination of a single V. cholerae strain throughout Iran in 2005.
Background:Pertussis is a respiratory and contagious disease which is mostly caused by Bordetella pertussis and B. parapertussis. It usually spreads from person to personduring the incubation or catarrhal phase of the disease. Despite of large-scale vaccination, whooping cough is still an endemic disease with several outbreaks.Objectives:The aim of this study was to determine the prevalence of pertussis and identify its causative agents, B. pertussis or B. parapertussis, from specimens collected from Iranian patients from 2004 to 2008.Patients and Methods:Nasopharyngeal swab samples from 347 suspected pertussis cases were collected from 18 provinces of Iran. The patients were in different age groups and were either unvaccinated or vaccinated for pertussis with whole cell vaccine (WCV). Bacterial culture, agglutination tests and quantitative PCR (qPCR) targeting IS481 and IS1001 for B. pertussis and B. parapertussis were done for every specimen, respectively.Results:The results showed that seven nasopharyngeal swab samples (2%) were positive for B. pertussis (1.7%) and B. parapertussis (0.3%) by culture and agglutination test and 30 patients had positive qPCR test results (9%).Conclusions:Despite the fact that bacterial culture is the golden standard for the detection of B. pertussis, direct detection of bacteria from nasopharyngeal specimens can be performed by a rapid qPCR assay. In this study, high percentage of positive qPCR cases may indicate that the patients might have recovered from pertussis following antibiotic treatment before samples were collected. Rapid detection by qPCR could be important for immediate diagnosis and treatment of patients with pertussis.
Background and Objectives: Cholera disease remains an important global health problem affecting 3-5 million subjects worldwide. Outer membrane vesicles (OMVs) have been found in a variety of Gram-negative bacteria and act as protective transport vesicles. The aim of this study was to evaluate Immune responses against Vibrio cholerae O1 El Tor clinical strain OMV and compare it with killed whole cell (KWC), complex of (KWC-OMV) as well as the internationally licensed oral cholera vaccine, Dukoral, in serum and intestinal secretions of mice. Materials and Methods: OMVs were prepared by using modified detergent-centrifugation procedure from V. cholerae O1 El Tor clinical strain from 2005 outbreak. The ultrastructure and content of OMVs were investigated via the Scanning Elec- tron Microscopy (SEM) and SDS-PAGE analysis. Three doses of oral immunization were adjusted and total IgG and IgA in serum and intestinal secretion were measured by enzyme-linked immunosorbent assay (ELISA). Results: Extracted OMVs from the V. cholerae were spherical vesicles with a size ranging from 10 to 300 nm. OMV-im- munized mice showed an increased level of total IgG and IgA both in serum and intestinal secretion when compared to the negative controls. Also, there existed a higher level of secretory IgA than the total IgG, suggesting the most of protection against V. cholerae colonization provided by sIgA. Conclusion: Our findings revealed that oral immunization with V. cholerae OMVs might induce a long-term immunity, es- pecially when administered in combination with KWC. This study tested the adjuvant activity of OMVs and may be useful in future nano vaccine research.
Even though cholera is easily prevented and treated, it remains life-threatening in Iran , this study indicated the need for continuous recording of antibiotic resistance and status of hlyA production Vibrio cholerae El tor at national level. Background: Cholera is a severe diarrheal illness caused by Vibrio cholerae, which produces a virulence factor named El Tor hemolysin encoded by the hlyA gene. Objectives: This study meant to analyze the phenotypic characteristics and presence of hlyA gene in V. cholerae isolated from patients in Iran. The hlyA gene which codes for hemolysin, plays an essential role in manifestation of cholera ,and could be used to diagnose pathogenic V. cholerae El Tor O1 strains. Patients and Methods: One hundred stool samples from the patients with cholera during 2002-2003 were collected from Tehran, Kashan, Kermanshah and Ahvaz cities, which were subject to diagnostic tests. Serotyping, and antibiotic susceptibility tests were applied and polymerase chain reaction (PCR) was also used to detect the hlyA gene. Results: The group specific antisera identified the isolates as Ogawa, Inaba, Hikojima and NAG (Non-agglutinable) in 74%, 3%, 0% and 23% of the isolates, respectively. Antibiotic susceptibility test showed that all of the strains were sensitive to ciprofloxacin, gentamycin and doxycyclin but the isolates showed resistance to sulfamethoxazole/trimethoprim (74%), erythromycin (64%) and tetracycline (50%). V. cholerae El Tor isolates were 100% positive for hlyA gene, but hemolysis phenotype characteristics were found in 95% of the cases. Conclusions:The results indicated that Ogawa serotype was identified as the dominant serotype which revealed multiple antibiotic resistances to sulfamethoxazole/trimethoprim, oxytetracycline, erythromycin, tetracycline and chloramphenicol. The presence of hlyA gene in nonhemolytic strains of V. cholerae O1 biotype El Tor indicated that some factors prevent expression of the hemolysin gene.
In this study, antimicrobial susceptibility test and genetic typing were used to characterize 15 Salmonella enterica serotype Typhi (S. Typhi) isolates recovered from sporadic cases of typhoid fever in Tehran, Iran during 2004. Antimicrobial susceptibility test showed that all isolates were susceptible to 20 antimicrobials examined in this study. Analysis of insertion elements showed that 2 IS200 types with 10 and 11 copies were present. 11 of the 15 isolates were found to possess 10 IS200 elements residing on fragments from 23 to 2.3 kb. Comparison of the RiboPrinter (automated ribotyping) patterns of S. Typhi showed that 60% (9/15) of the isolates belonged to a single ribotype. PCR based random amplified polymorphic DNA analysis (RAPD) and enterobacterial repetitive intergenic consensus (ERIC) and pulsed-field gel electrophresis (PFGE) were also performed. ERIC and RAPD-PCR method showed 2 and 3 genotyping patterns amongst the isolates, respectively. The PFGE typing was carried out by using XbaI restriction enzyme, and 7 restriction patterns were observed. Overall, the molecular typing methods applied in this study showed that the isolated S. Typhi populations were highly polyclonal as shown by PFGE.
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