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Showing 4 results for Salmonella

Abobakr Moradi, Dr Ali Karami, Dr Ali Hagh Nazari, Zeinab Ahmadi, Dr Rahim Soroori Zanjani, Seyedeh Mehri Javadi,
Volume 17, Issue 67 (8-2009)
Abstract

Background and Objective: There are several techniques for the diagnosing of salmonella infectious. Several molecular methods such as PCR and hybridization assay have recently been used for the detection of this bacterium. However, these methods require precision instruments for amplification and complex procedures, which are the major obstacles to the widespread use of these methods in relatively small scale clinical laboratories, clinics and the filed laboratories. Recently, a new, rapid and sensitive technique called loop-mediated isothermal amplification (LAMP) was developed. Materials and Methods: In this study we used 7 different strains of salmonella to compare the PCR with LAMP method. For PCR test we used thermocycler, but The LAMP reaction can be conducted under isothermal conditions by using only one type of enzyme and four primers recognizing six distinct regions. The most important merit of this method is that no denaturation of the DNA template is required, so, technique is simple and no need to thermocycler machine and several temperatures cycles. Results: Conventional PCR method for the detection of Salmonella with standard thermocylcer takes 3 hrs but, with LAMP method we were able to amplify and detect the salmonella in very simple thermal block made in IRAN. After Optimization of the process it was possible to rapidly detect and identify Salmonella typhi bacteria within 90 minutes. This method was also 100 times more sensitive comparing to the PCR method. Conclusion: According to the results, comparing LAMP isothermal amplification method for detection and identification of Salmonella with conventional PCR we have been able to determine the simplicity, speed (3 times) and the superior sensitivity(100 times) of the LAMP to PCR method. This Method is more simple, faster and cheaper (10 times). Another advantage is independence to cycle's temperature and thermo-cycling and replacement with one thermo block which is very simple, inexpensive and made in inside the country.


Reza Ranjbar, Meysam Sarshar, Nour Khoda Sadeghifard,
Volume 20, Issue 81 (9-2012)
Abstract

Background and Objective: Salmonella spp. are enteric pathogens with a worldwide distribution comprising a large number of serovars characterized by different hosts and distribution. Among Salmonella spp., the number of infections and diseases caused by the serotype Salmonella enterica serovar Infantis started to increase significantly in the last decade. The aim of this study was to investigate the genetic diversity of the clinical stains of Salmonella enterica serovar Infantis isolated in Tehran, Iran by using the Ribotyping method. Materials and Methods: In this descriptive study from November 2007 to December 2010, clinical samples, collected from different hospitals in Tehran, were investigated for detection of Salmonella enterica serovar Infantis. Bacterial isolation and identification was achieved through biochemical and bacteriological methods. The Ribotyping technique was applied for the molecular typing of the strain of Salmonella enterica serovar Infantis. Results: Out of the 26 Salmonella serogroup C samples isolated in this study, 19 strains (73%) belonged to Salmonella enterica serotype Infantis. Ribotyping results divided Salmonella enterica serovar Infantis stains into 9 clusters (1c to 9c). The majority (7) of the strains belonged to cluster 1c. Conclusion: The results obtained from the Ribotyping patterns indicate that Salmonella enterica serovar Infantis strains, circulating among the patients in Tehran, belong to a diverse number of clones. Moreover, our data show that Ribotyping is an appropriate method for the molecular typing of Salmonella enterica serovar Infantis strains.


Narges Safari Foroshani, Ali Karami, Fatemeh Pourali, Akram Aeidi,
Volume 21, Issue 85 (4-2013)
Abstract

Background and Objectives: Salmonella typhi, Bacillus anthracis, and Yersinia pestis are potent human pathogenic bacteria that cause typhoid fever, anthrax, and plague,respectively. The classic microbiological methods for detection and identification of these agents are laborious and time consuming.Therefore, developing an accurate method for rapid detection of these potent human pathogens is important, especially due to their potential use as Bioterrorism agents. We have designed a new rapid molecular detection method by using multiplex PCR in a single reaction. Materials and Methods: PCR was carried out using specific primers for the virulence factor of S. typhi, hp and invA, B. anthracis virulence factors, chr and pa, and Y. pestis toxin, pla. The primer sets were tested for cross reactivity with individual DNA strains and mixtures in a multiplex PCR format. For determination of sensitivity of the method, we used colony counting and genomic DNA dilution. Results: Theresult with standard strains revealed that primers are specific for each strain as they have successfully amplified the desire products. The sensitivity analysis showed a 1-10 CFU and 1-10 copies of the genomes. In this respect, the accuracy of the result was checked by sequencing of the 1083-bp PCR products, except for anthrax, which was determined by enzymatic digestion. Conclusion: We have found a rapid, specific, sensitive, and a non-expensive molecular method for detection of three potent human pathogens. This finding provide an efficient diagnostic tool to determine these bacteria in bioterrorism samples.


Mohammad Reza Esmaeil Zadeh, Mohammad Kazem Sharifi Yazdi, Zahra Rajabi, Farzaneh Amin Harati, Farhad Nikkhahi, Sara Sharifi Yazdi, Gholamreza Hassanpour, Alireza Monadi Sefidan, Mohammad Mehdi Soltan Dallal,
Volume 30, Issue 139 (1-2022)
Abstract

Background and Objective: Phage therapy could be used as an alternative method to antibiotic treatments. The purpose of this study was to evaluate the antibacterial activities of isolated lytic bacteriophage against ciprofloxacin-resistant strain of Salmonella infanits in vitro conditions.
Materials and Methods: The standard strain of Salmonella infantis  and its specific bacteriophage was isolated by soft agar method. Phage susceptibility to heat and pH was evaluated by the Double-Layer Agar method.  In vitro assay was carried out on human epithelial type 2 (HEp-2) cells to investigate the effect of bacteriophage on the cytotoxic and invasion of Salmonella infantis to human epithelial cells.
Results: Head and tail morphology of bacteriophages against Salmonella infantis were identified by transmission electron microscopy and assigned to the Myoviridae family. The results of the double-layer agar assay showed that the titer of bacteriophages was 1.8×107 PFU/ml. bacteriophage was stable at 4 ֯C and the best quantification of bacteriophage was determined at pH=8. The isolated bacteriophage was specific for Salmonella infantis and had no lytic activity against other pathogenic bacteria. In the evaluation of the binding and invasion of Salmonella infantis to the HEp-2 cell line, as expected, the lytic activity of specific bacteriophage was observed following inoculation.
Conclusion: Additional studies are needed for better understanding of the interaction between phage, microorganisms and human host before applying phage therapy on a large scale.



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