Application of Reverse Transcriptase -PCR (RT-PCR) for rapid detection of viable Escherichia coli in drinking water samples

Application of Reverse Transcriptase -PCR (RT-PCR) for rapid detection of viable Escherichia coli in drinking water samples


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نویسندگان: حمید ابطحی , احسان اله غزنوی راد , محمدجواد قنادزاده , محمود کریمی , ندا مولایی

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نشریه: ---- عنوان نشریه در لیست موجود نیست ----, 24,13,1 - 6,2015

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کد مقاله 2872
عنوان فارسی مقاله Application of Reverse Transcriptase -PCR (RT-PCR) for rapid detection of viable Escherichia coli in drinking water samples
عنوان لاتین مقاله Application of Reverse Transcriptase -PCR (RT-PCR) for rapid detection of viable Escherichia coli in drinking water samples
نوع مقاله بر حسب نگارش پژوهشی اصیل
مقاله برحسب نمایه ISI
IF 0.5
عنوان نشریه Journal of environmental health science & engineering
نوع نشریه علمی پژوهشی
شماره نشریه 24
دوره 13
صفحه شروع و پایان در نشریه 1 - 6
سال انتشار/ ارائه شمسی 1393
سال انتشار/ارائه میلادی 2015
آدرس لینک مقاله/ همایش در شبکه اینترنت http://apps.webofknowledge.com/full_record.do?product=UA&search_mode=GeneralSearch&qid=1&SID=N2Wtx9p7FD65gNobXVe&page=1&doc=1
ISSN 2052-336X
DOI DOI: 10.1186/s40201-015-0177-z
آدرس علمی (Affiliation) نویسنده متقاضی Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran

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چکیده انگلیسیApplication of Reverse Transcriptase -PCR (RT-PCR) for rapid detection of viable Escherichia coli in drinking water samples Molaee N1, Abtahi H2, Ghannadzadeh MJ3, Karimi M4, Ghaznavi-Rad E5 • 1Department of Microbiology and Immunology, Arak University of Medical sciences, Arak, Iran. • 2Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran. • 3Department of Environmental Health, Faculty of Health, Arak University of Medical Sciences, Arak, Iran. • 4Department of Medical Microbiology and Immunology, Arak University of Medical sciences, Arak, Iran. • 5Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran ; Department of Medical Microbiology and Immunology, Faculty of Medicine, Arak University of Medical Sciences, Arak, Iran. Abstract BACKGROUND: Polymerase chain reaction (PCR) is preferred to other methods for detecting Escherichia coli (E. coli) in water in terms of speed, accuracy and efficiency. False positive result is considered as the major disadvantages of PCR. For this reason, reverse transcriptase-polymerase chain reaction (RT-PCR) can be used to solve this problem. The aim of present study was to determine the efficiency of RT- PCR forrapid detection of viable Escherichia coli in drinking water samples and enhance its sensitivity through application of different filter membranes. MATERIALS AND METHODS: Specific primers were designed for 16S rRNA and elongation Factor II genes. Different concentrations of bacteria were passed through FHLP and HAWP filters. Then, RT-PCR was performed using 16srRNA and EF -Tu primers. Contamination of 10 wells was determined by RT-PCR in Arak city. To evaluate RT-PCR efficiency, the results were compared with most probable number (MPN) method. RESULTS: RT-PCR is able to detect bacteria in different concentrations. Application of EF II primers reduced false positive results compared to 16S rRNA primers. The FHLP hydrophobic filters have higher ability to absorb bacteria compared with HAWB hydrophilic filters. So the use of hydrophobic filters will increase the sensitivity of RT-PCR. CONCLUSION: RT-PCR shows a higher sensitivity compared to conventional water contamination detection method. Unlike PCR, RT-PCR does not lead to false positive results. The use of EF-Tu primers can reduce the incidence of false positive results. Furthermore, hydrophobic filters have a higher ability to absorb bacteria compared to hydrophilic filters. KEYWORDS: Coliforms; Drinking water; Reverse transcriptase
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نویسنده نفر چندم مقاله
حمید ابطحیدوم
احسان اله غزنوی رادپنجم و مسئول
محمدجواد قنادزادهسوم
محمود کریمیچهارم
ندا مولاییاول

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