Detection of Chicken-Derived Components in Livestock and Poultry Meat by Fluorescent PCR Assay
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Abstract:
In order to establish a fluorescent quantitative PCR method for detecting the components of chicken origin in livestock and poultry meat, the mitochondrial DNA 16S rRNA gene sequences of 9 animals including chicken, duck, goose, pig, cattle, sheep, rabbit, pigeon and quail were used as target sites. The chicken-specific primers were screened through comparative analysis design, and specificity amplification was performed using 9 kinds of animal muscle DNA as templates to establish the conditions for fluorescent quantitative PCR amplification. In the meantime, the chicken DNA template concentration was serially diluted 10-fold to 108 times. The sensitivity of the established fluorescent PCR method was tested Then the samples randomly collected from the market were analysed using this established method. The results showed that, the designed chicken primer had a typical amplification curve only for the chicken DNA template (Ct value as 22.11), but no amplification for other animal DNA templates, thus the method exhibited high specificity. When the chicken DNA template dilution reached 104 times, that is, the DNA concentration was 17.5 pg/μL, there was still a typical amplification curve with Ct value as 30.37, thus the method had relatively high sensitivity. Analyses of the samples randomly collected from the market circulation revealed that all the collected samples were qualified. It is clear that the method for detecting the components of chicken origin in livestock and poultry meat is fast and accurate with strong practicality.
