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A molecular beacon-based real time NASBA assay for detection of Listeria monocytogenes in food products : role of target mRNA secondary structure on NASBA designNadal, Anna ...A molecular beacon-based real-time NASBA (QNASBA) assay for detection and identification of Listeria monocytogenes has been developed. A correlation between targeting highly accessible mRNA sequences ... and QNASBA efficiency and sensitivity was demonstrated. The assay targets a sequence from the mRNA transcript of the hly gene which is specific for this bacterium; and includes an internal amplification control to disclose failure of the reaction. It wasfully selective and consistently detected down to 100 target molecules and40 L. monocytogenes exponentially growing cells per reaction. In addition, it was capable of accurate quantification of target RNA molecules independently of the presence of DNA in the sample. In combination with a short RNase treatment prior to nucleic acids extraction our QNASBA specifically detected viable L. monocytogenes cells. It was successfully applied to rapid detection of this pathogen in meat and salmon products, and is therefore a useful tool for the study of L. monocytogenes in food samples. We finally discuss considerations of target secondary structure with regard todevelopment of NASBA assays.Source: Journal of microbiological methods. - ISSN 0167-7012 (Vol. 68, issue 3, 2007, str. 623-632)Type of material - article, component partPublish date - 2007Language - englishCOBISS.SI-ID - 28777945
Author
Nadal, Anna |
Coll Rius, Anna |
Nigel, Cook |
Pla, Maria
Topics
Molecular Probes |
Animals |
Base Sequence |
Dna Primers |
Listeria Monocytogenes |
Genetics |
Growth & Development |
Isolation & Purification |
Meat Products |
Microbiology |
Molecular Sequence Data |
Nucleic Acid Conformation |
Rna, Bacterial |
Genetics |
Chemistry |
Isolation & Purification |
Rna, Messenger |
Chemistry |
Genetics |
Isolation & Purification |
Salmon |
Microbiology |
Molekularne sonde |
RNA, prenašalna |
Listeria monocytogenes |
Molekulsko zaporedje, podatki |
Živali |
Nukleinska kislina, konformacija |
Baze, zaporedje |
DNA začetki |
RNA bakterijska |
Mesni izdelki |
Losos
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Nadal, Anna | ![]() |
Coll Rius, Anna | 34130 |
Nigel, Cook | ![]() |
Pla, Maria | ![]() |
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