Evaluation and comparison of SYBR Green I Real-Time PCR and TaqMan Real-Time PCR methods for quantitative assay of Listeria monocytogenes in nutrient broth and milk
Data(s) |
09/03/2012
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Resumo |
Specific traditional plate count method and real-time PCR systems based on SYBR Green I and TaqMan technologies using a specific primer pair and probe for amplification of iap-gene were used for quantitative assay of Listeria monocytogenes in seven decimal serial dilution series of nutrient broth and milk samples containing 1.58 to 1.58×107 cfu /ml and the real-time PCR methods were compared with the plate count method with respect to accuracy and sensitivity. In this study, the plate count method was performed using surface-plating of 0.1 ml of each sample on Palcam Agar. The lowest detectable level for this method was 1.58×10 cfu/ml for both nutrient broth and milk samples. Using purified DNA as a template for generation of standard curves, as few as four copies of the iap-gene could be detected per reaction with both real-time PCR assays, indicating that they were highly sensitive. When these real-time PCR assays were applied to quantification of L. monocytogenes in decimal serial dilution series of nutrient broth and milk samples, 3.16×10 to 3.16×105 copies per reaction (equals to 1.58×103 to 1.58×107 cfu/ml L. monocytogenes) were detectable. As logarithmic cycles, for Plate Count and both molecular assays, the quantitative results of the detectable steps were similar to the inoculation levels. |
Formato |
text |
Identificador |
http://centaur.reading.ac.uk/28742/1/25%20Dadkhah%20et%20al%202012.pdf Karatzas, K. A. G. <http://centaur.reading.ac.uk/view/creators/90004634.html> (2012) Evaluation and comparison of SYBR Green I Real-Time PCR and TaqMan Real-Time PCR methods for quantitative assay of Listeria monocytogenes in nutrient broth and milk. African Journal of Microbiology Research, 6 (9). pp. 1908-1917. ISSN 1996-0808 doi: 10.5897/AJMR11.889 <http://dx.doi.org/10.5897/AJMR11.889> |
Idioma(s) |
en |
Publicador |
Academic Journals |
Relação |
http://centaur.reading.ac.uk/28742/ creatorInternal Karatzas, Kimon Andreas G. 10.5897/AJMR11.889 |
Tipo |
Article PeerReviewed |