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Evaluation of LAMP (Loop-mediated isothermal DNA amplification) for Molecular Detection of Salmonella
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Ali Karami *   |
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Abstract: (14229 Views) |
| Introduction: Salmonella Bacterium is not only a causative factor of typhoid fever, enterocolitis, and salmonellosis, but it is also a zoonotic infection. This bacterium is a major health problem throughout the world, and is especially prevalent in developing countries. Therefore, rapid diagnosis of salmonella can prevent its outbreak. Different techniques are used for the diagnosis of Salmonella bacteria, such as culture, biochemical, serological, ELISA, Widal, immunofluorescence and molecular methods like PCR and Real time PCR,all of which are difficult, time-consuming, and expensive. Thus, our study was designed to evaluate the LAMP (Loop-mediated isothermal DNA amplification method) for detection of salmonella bacteria. Materials & Methods: In this study, we examined 7 different strains of salmonella. The DNA was extracted by standard methods and amplified with specific primers by PCR and set of primers for LAMP in single temperature in very simple thermal block made in Iran. The amplified products were detected by gel electrophoresis and LAMP products were visualized by their turbidity with naked eye. Findings: Conventional PCR method for detection of Salmonella needs standard thermocylcer and takes 3 hours, but using LAMP method, we were able to amplify and detect salmonella in simple thermoblock, taking much less time. After optimization of the process, it was, soon, possible to detect and identify Salmonella typhi bacteria within 90 minutes. This method is also 10 times more sensitive than that of the PCR. Discussion & Conclusion: According to the results, comparing LAMP method for detection and identification of Salmonella with conventional PCR, we have been able to determine the simplicity, speed and the superior sensitivity of the LAMP method. This Method is more simple, faster and cheaper. Non-dependence of cycle's temperature and thermo-cycling and replacement with one thermo block which is very simple, inexpensive and made inside the country, can be considered another advantage of the LAMP method. |
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| Keywords: Salmonella, molecular detection, PCR, DN, LAMP |
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Full-Text [PDF 461 kb]
(4271 Downloads)
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Type of Study: Research |
Subject:
General
Received: 2010/02/9 | Accepted: 2015/04/20 | Published: 2015/04/20
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