Babak barati; Shahram Nazarian; Mehdi Shirazi; Nasibeh Ghorbani; Mojtaba Saadati; Mohammadbagher Salehi; Hadi Shirzad
Volume 16, Issue 4 , January and February 2010, , Pages 221-227
Abstract
Background and Purpose: Typhoid fever, a disease caused by Salmonella typhi, is still one of the most important infectious diseases across the world. Different methods such as biochemical and Elisa methods are used for detection of this bacterium, which produce false responses in addition to being time-consuming ...
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Background and Purpose: Typhoid fever, a disease caused by Salmonella typhi, is still one of the most important infectious diseases across the world. Different methods such as biochemical and Elisa methods are used for detection of this bacterium, which produce false responses in addition to being time-consuming and expensive. Therefore, the present research was conducted to detect Salmonella typhi by PCR method which is rapid, inexpensive and specific. Materials and methods: In this descriptive study which was conducted via diagnostic method, polymerase chain reaction (PCR) assay was developed for detection of Salmonella typhi. This strain had formerly been confirmed by biochemical methods. For detection by PCR, one primer pair was designed, being specific to ViaB gene. The PCR product was digested by restricted enzyme. For specificity of assay, 6 different strains were used as control negative and for sensitivity of PCR reaction, serial dilution of bacteria was used. Results: The PCR product of Salmonella typhi was 530 bp which were then confirmed by digestion enzymes. In testing the specificity of the assay, Salmonella typhimorium, Shigella flexneri, E. coli, Clostridium botulinum, Staphylococcus aureus and Bacillus subtilis were used as negative control, and did not yield a PCR product. The sensitivity of this method was estimated to be about 50 CFU/ml. Conclusion: The results of this study suggest that detection of ViaB gene with PCR method can be used for diagnossis of Salmonella typhi in clinical samples as a rapid, inexpensive, specific and highly sensitive method.