转基因溶菌酶羊hLZ-J1品系特异性数字PCR方法及样本中痕量转基因成分检测

人溶菌酶是一种天然抗生素，具有抗细菌、真菌，抗炎症，预防感染等功能。转基因溶菌酶山羊可以在乳腺中特异性表达，从而可以从羊乳中规模化制备溶菌酶。本研究旨在建立一种适用于痕量样本中转基因溶菌酶羊成分定量检测的品系特异性数字PCR方法，并利用该方法对山羊的血液、羊奶、粪便和环境土壤样本等进行检测，用于对转基因溶菌酶山羊成分的溯源。建立的数字PCR方法具有高特异性、灵敏度、准确性，检测限和定量限低至15个拷贝每个反应，可以对羊血、羊乳等制品进行准确的定量检测。研究结果还表明该方法可以准确测定粪便和环境土壤样本中的痕量转基因溶菌酶羊成分，可进一步用于转基因动物外源基因的水平转移检测及环境风险评估。

Development of an Event-Specific Droplet Digital PCR Assay for Quantification of the Transgene DNAs in Trace Samples of GM Goat hLZ-J1

Human lysozyme (hLZ) is a natural antibiotic,functioning to resist bacteria,fungi,inflammation and infection.Transgenic human lysozyme goat can express high levels of human lysozyme in milk for large scale production of lysozyme.Herein,we developed an event-specific droplet digital PCR (ddPCR) method for quantitative detection of transgenic hLZ goat and its derivates.This method presents high specificity,sensitivity and accuracy in goat blood and goat milk samples analysis.The limit of detection and limit of quantification were determined with 15 copies per reaction.Furthermore,this method was used to trace the transgenic hLZ goat content in goat feces and the living environmental soil samples,indicating that the developed ddPCR method can be used for monitoring horizontal gene transfer and environmental risk assessment of transgenic hLZ goat.