Rapid Detection Technique of Duck-Derived Components in Beef and Sheep Products by LAMP

Abstract:

A pair of specific PCR primers was selected based on the principle of animal species specificity in order to establish a duck meat products source components on-site rapid detection technology. The fragment was obtained by the specific primers only in duck samples. On the basis of this fragment，the ring mediated isothermal amplification（LAMP） primers were designed and the primers could detect duck source component specifically. By conditions optimization experiments，the most optimal amplification system for LAMP（25 ?滋L） as follow：concentration ratio of inside primer and outside primer was 1∶4，temperature was 63.5 ℃，MgSO4 was 5 mM/μL，dNTPs was 1.75 mM/μL，Bst DNA polymerase was 0.4 U/μL，betainewas 0.25 μmol/μL，the LAMP reaction time was 30 min. Under the optimized conditions，the detection sensitivity of duck DNA was 10 pg/uL. The detection sensitivity of component proportion for beef or mutton in the duck source was 0.000 1%. 21 samples from markets of Shanghai Minhang district were detected using this method and 5 samples contained duck source. The results showed LAMP was a sensitive rapid technology. It provides a new method to realize on-site detection of duck source components for meat products.