| Autor: |
Mahmoudzadeh, Maryam, Hosseini, Hedayat, Hedayati, Mehdi, Mousavi Khanghah, Amin, Djalma Chaves, Rafael, Azizkhani, Maryam |
| Předmět: |
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| Zdroj: |
Journal of Food Safety; May2016, Vol. 36 Issue 2, p220-226, 7p |
| Abstrakt: |
Ingredients and complexity of food matrices are inhibitory factors for gene expression studies of bacterial pathogens directly from food matrix. The aim of this study was to develop a simple and fast method for RNA extraction from ground beef matrix as well as stx genes expression of E scherichia coli O157: H7 by relative quantitative real-time reverse transcription-polymerase chain reaction ( RT-PCR). Inoculated ground beef samples were kept 7 days in refrigerated storage (4 ± 1 C) and bacterial RNA was extracted according to the described method at 0, 2, 5 and 7 days after storage. Results indicate that refrigerated storage temperature decreased gene expression of stx1A (−3.35 relative changes on day 7) and stx2A (−2.9 relative changes on day 7) in ground beef matrix. In the current study, a method requiring minimal equipment and limited interference for RNA extraction from bacteria associated with a meat matrix was evaluated (developed, invested, etc.). Results of the study suggest that the quantitative real-time RT-PCR method could be used as a reliable and sensitive method for gene expression studies. Practical Applications Polymerase chain reaction ( PCR), known as a sensitive and specific method, was extensively used for the detection of genes involved in stress and virulence of bacteria. In this way, real-time PCR shows a good sensitivity and diversity for detection and gene expression studies of pathogens. This technique frequently has been used for the study of bacterial behavior in broth or food-like broth models. Complexity of food matrix and presence of proteins, and also other PCR-inhibiting factors, restricted gene expression studies in food matrix. There has been great interest in the survival and pathogenicity of E scherichia coli O157:H7 because it is a great concern of ground meat industry. Shiga toxins ( Stx1 and Stx2) as manifest characteristics of E . coli O157:H7 are known as significant virulence factors in the pathogenicity of bacterium so its behavior and gene expression profile in meat matrix and during cold chain were evaluated. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
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