Citation
Hayati M., . and Sudjadi, . and Rohman A, . Analysis of Salmonella enteritidis in chicken meat and egg by real timepolymerase chain reaction. pp. 2689-2693.
Abstract
The objective of this study was to develop analytical method for analysis of Salmonella enteritidis contamination in chicken meat and eggs using real time polymerase chain reaction (real-time PCR) method. A pair of primers have been designed using NCBI website and is proven to amplify Sdf1 genes a specific gene of S. enteritidis. The study involved preenrichment of bacterial culture of S. enteritidis DNA isolation validation real-time PCR method and application of validated method to analyze S. enteritidis in the commercial product. Isolation of DNA was performed using phenol-chloroform method. A range of temperature (51.0-62.2C) was examined to obtain optimum annealing temperature. The temperature of 60.2C was chosen based on melting peak profile. Specificity test performed toward four bacterial DNA (S. enteridis S. typhimurius Escherichia coli and Listeria monocytogenes) and chicken DNA showed that this primer specifically amplify S. enteritidis with melting peak point at 77.5-78.0C. Sensitivity test resulted the limit of detection at 12.5 pg/L for S. Enteritidis DNA. The results showed that no amplification peak in commercial products indicating that the tested commercial products do not contain S. enteritidis.
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Abstract
The objective of this study was to develop analytical method for analysis of Salmonella enteritidis contamination in chicken meat and eggs using real time polymerase chain reaction (real-time PCR) method. A pair of primers have been designed using NCBI website and is proven to amplify Sdf1 genes a specific gene of S. enteritidis. The study involved preenrichment of bacterial culture of S. enteritidis DNA isolation validation real-time PCR method and application of validated method to analyze S. enteritidis in the commercial product. Isolation of DNA was performed using phenol-chloroform method. A range of temperature (51.0-62.2C) was examined to obtain optimum annealing temperature. The temperature of 60.2C was chosen based on melting peak profile. Specificity test performed toward four bacterial DNA (S. enteridis S. typhimurius Escherichia coli and Listeria monocytogenes) and chicken DNA showed that this primer specifically amplify S. enteritidis with melting peak point at 77.5-78.0C. Sensitivity test resulted the limit of detection at 12.5 pg/L for S. Enteritidis DNA. The results showed that no amplification peak in commercial products indicating that the tested commercial products do not contain S. enteritidis.
Additional Metadata
| Item Type: | Article |
|---|---|
| AGROVOC Term: | Salmonella enteritidis |
| AGROVOC Term: | Gram negative bacteria |
| AGROVOC Term: | Salmonella typhimurium |
| AGROVOC Term: | Escherichia coli |
| AGROVOC Term: | Listeria monocytogenes |
| AGROVOC Term: | Chicken meat |
| AGROVOC Term: | Contamination |
| AGROVOC Term: | analysis |
| AGROVOC Term: | Identification |
| AGROVOC Term: | Isolation |
| Depositing User: | Ms. Suzila Mohamad Kasim |
| Last Modified: | 24 Apr 2025 06:28 |
| URI: | http://webagris.upm.edu.my/id/eprint/23936 |
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