肉類食品真?zhèn)舞b定是目前食品安全檢測領(lǐng)域的重點(diǎn)研究內(nèi)容之一。為準(zhǔn)確定量檢測食品中的雞源性成分，根據(jù)現(xiàn)行檢驗(yàn)檢疫標(biāo)準(zhǔn)（SN/T 2727—2010）合成引物及探針，并拓展了特異性、靈敏度、檢出限和定量分析等試驗(yàn)。結(jié)果顯示：特異性試驗(yàn)中，僅雞成分檢測出現(xiàn)特異性擴(kuò)增，其他動物組織均未出現(xiàn)特異性擴(kuò)增；梯度稀釋雞源性DNA模板原液進(jìn)行靈敏度評價，發(fā)現(xiàn)引物和探針靈敏度較高，可以檢測到100 fg級的雞組分模板DNA，且建立的標(biāo)準(zhǔn)曲線具有良好的線性關(guān)系，R2達(dá)0.99以上，定量準(zhǔn)確；加工制品中的驗(yàn)證試驗(yàn)結(jié)果顯示，合成的引物及探針具有良好的適用性和定量檢測能力。以上研究結(jié)果均說明，此引物和探針適用于市場食品中雞源性成分的定性、定量檢測。本研究為今后TaqMan實(shí)時熒光PCR法不同動物源性引物和探針的自主研發(fā)奠定了基礎(chǔ)。
Quantitative Detection of Chicken-derived Components in
Food Based on Real-time PCR
At present，the identification of meat products is emphatically studied in the field of food safety. In order to establish a quantitative detection method for chicken-derived components in food，primers and probes were synthesized according to current standard of inspection and quarantine（SN/T 2727—2010），and specificity test，sensitivity test，detection limit and quantitative analysis were expanded. The results showed that the specific amplification was found only in chicken-derived components instead of other animal tissues；gradient dilution of chicken-derived DNA template were used to evaluate the sensitivity，and it was found that the sensitivity of primers and probe was high，which could detect the DNA template at 100 fg level，and a good linear relationship was observed in the standard curve，R2 could reach above 0.99 with an accurate quantitative detection. For the processed products，the synthesized primers and probes were with good applicability and quantitative detection capacity. It was concluded that the primers and probes were suitable for qualitative and quantitative detection of chicken-derived components in food markets. A foundation was laid for the independent development of different animal-derived primers and probes of TaqMan real-time PCR in the future.
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