year 3, Issue 2 And 3 (12-2009)                   Iran J Med Microbiol 2009, 3(2 And 3): 9-14 | Back to browse issues page

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Najafi A, Qorbanalizadgan M, Tavakoli H, Ahmadi A. Rapid ِِ Diagnosis of Listeria monocytogenesby PCR Method with hlyAgene . Iran J Med Microbiol 2009; 3 (2 and 3) :9-14
URL: http://ijmm.ir/article-1-151-en.html
1- Molecular Biology Research Center, Baghiatollah University of Medical Sciences, Tehran, Iran
2- Nutrition Department, Public Health Research Center, Baghiatollah University of Medical Sciences, Tehran, Iran. , h.tavakoli1344@yahoo.com
Abstract:   (19228 Views)
Background and Objectives: Currently the conventional method for the diagnosis of Listeria monocytogenes is culture method. However, the isolation rate is reduced in culture due to animals' food which is contains antibiotics. Besides, culture method is time consuming and takes more than 36 hrs. So more precise and rapid technique is needed. The aim of this study was application of PCR technique based on hlyA gene fragment for rapid detection of Listeria monocytogenes in clinical samples.
Materials and Methods: hlyA gene was selected as a specific target sequence for detection of Listeria monocytogenes. Listeria monocytogenes PTCC1163 was used as a standard organism for optimization experiments. A range of bacterial pathogens was used for specificity test including Escherichia coli: ATCC 35218 Salmonella typhi PTCC: 1609. Phenol – chloroform method was used for DNA extraction. PCR technique was performed as per standard protocol. Amplified product was detected by 1% gel agarose electrophoresis, stained by ethidiome bromide.
Results: Provided data confirmed amplification of expected product. Specificity test proved no cross reaction with tested organisms. Sensitivity test detected 500fg Listeria monocytogenesDNA as a final detection limit.
Conclusion: Optimized experiment confirmed that the applied PCR protocol is quite fast with high sensitivity and specificity performing in less than three hours. Application of this method in clinical laboratories can help the rapid diagnosis of Listeria monocytogenesin samples.
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Type of Study: Original Research Article | Subject: Medical Bacteriology
Received: 2013/11/22 | Accepted: 2013/11/22 | ePublished: 2013/11/22

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