COMPARISON OF DNA EXTRACTION METHO DS TO DETECT SALMONELLA SPP. IN TAP WATER.
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| Title: | COMPARISON OF DNA EXTRACTION METHO DS TO DETECT SALMONELLA SPP. IN TAP WATER. |
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| Alternate Title: | PRI MER JAVA METOD OSA MITVE DNA ZA ODKRIVAN JE BAKTERI J IZ RO DU SALMONELLA VPITNIVODI. |
| Authors: | Ocepek, Matjaž1 matjaz.ocepek@vf.uni-lj.si, Pate, Mateja1, Kušar, Darja1, Hubad, Barbara2, Avberšek, Jana1, Logar, Katarina1, Lapanje, Aleš2, Zrimec, Alexis2 |
| Source: | Slovenian Veterinary Research / Slovenski Veterinarski Zbornik. 2011, Vol. 48 Issue 3/4, p93-98. 6p. |
| Document Type: | Article |
| Subjects: | Salmonella, Water pollution, Salmonella food poisoning, DNA, Drinking water |
| Author-Supplied Keywords: | DNA extraction food safety pathogen detection PCR water odkrivanje patogenov osamitev DNA PCR Salmonella varna hrana voda Language of Keywords: English; Slovenian |
| Abstract (English): | Bacteria of the genus Salmonella cause a global health problem related to contaminated foodstuffs and faecally- polluted water supplies. The aim of our work was to determine the efficiency and rapidness of Salmonella detection in spiked tap water samples by comparison of different methods for DNA extraction. Water samples were spiked with different Salmonella loads. Four DNA extraction methods were employed: foodproof™ Sample Preparation Kit I (Biotecon Diagnostics) and QIAamp® DNA Stool Mini Kit (Qiagen) for DNA extraction after an overnight enrichment step, and Adiapure® Water DNA Extraction and Purification Kit (Adiagene) and SmartHelix® Complex Samples DNA Extraction Kit (developed recently in our laboratory) for direct DNA extraction. The extracted DNA was subjected to PCR amplification using Salmonella genus-specific primer pair. The SmartHelix extraction kit proved to be more efficient than the Adiapure Water kit for direct detection of Salmonella DNA in water. The overnight enrichment step improved the detection of Salmonella spp. in samples spiked with low bacterial load, however it extended the analysis time. Results suggest that the newly developed SmartHelix extraction kit should be regarded as a considerable choice when rapid detection of pathogens in water samples is aimed for to prevent disease outbreaks and support safe food assurance. [ABSTRACT FROM AUTHOR] |
| Abstract (Slovenian): | Bakterije iz rodu Salmonella predstavljajo splošen zdravstveni problem, povezan z okuženimi živili in fekalno onesnaženo vodo. Namen našega dela je bil s primerjavo različnih metod osamitve DNA ugotoviti učinkovitost in hitrost odkrivanja salmonel v vzorcih pitne vode, ki smo jim predhodno dodali omenjene bakterije v različnih koncentracijah. Uporabili smo štiri različne metode osamitve DNA: komercialna kompleta foodproof™ Sample Preparation Kit I (Biotecon Diagnostics) in QIAamp® DNA Stool Mini Kit (Qiagen) za osamitev DNA po čeznočni obogatitvi ter kompleta Adiapure® Water DNA Extraction and Purification Kit (Adiagene) in SmartHelix® Complex Samples DNA Extraction Kit, ki smo ga nedavno razvili v našem laboratoriju, za direktno osamitev DNA. DNA smo uporabili za pomnoževanje v PCR reakciji s parom začetnih oligonukleotidov, ki je specifičen za rod Salmonella. Komplet SmartHelix se je za direktno odkrivanje DNA salmonel v vodi izkazal kot bolj učinkovit v primerjavi s kompletom Adiapure Water. Čeznočna obogatitev je izboljšala odkrivanje salmonel v vzorcih, ki smo jim dodali majhno koncentracijo bakterij, vendar pa je podaljšala čas analize. Rezultati kažejo na to, da bi morali na novo izdelani komplet SmartHelix upoštevati kot eno izmed možnih izbir, ko želimo v vzorcih vode patogene bakterije odkriti hitro z namenom preprečevanja izbruhov bolezni in zagotavljanja varne hrane. [ABSTRACT FROM AUTHOR] |
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| Author Affiliations: | 1Institute of Microbiology and Parasitology, Veterinary Faculty, Gerbičeva 60, 1115 Ljubljana 2Institute of Physical Biology, Toplarniška 19, 1000 Ljubljana, Slovenia |
| ISSN: | 1580-4003 |
| Accession Number: | 70472911 |
| Database: | Veterinary Source |
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