鹅源性成分PCR检测方法的建立

Establishment of PCR Detections for Goose-derived Components

[目的]建立一种检测鹅源性成分的PCR方法。[方法]以15种动物的DNA为模板,利用鹅的特异性引物进行PCR反应,PCR产物用1.4%的琼脂糖凝胶电泳检测并测序,检测引物的特异性和敏感性。[结果]通过PCR反应从鹅样品中扩增到了304 bp的目的片段,其他物种火鸡、鸭、鹌鹑、鸵鸟、鸽子、鸡、马、牛、羊、猪、狗、驴、猫、鱼和空白对照则无目的片段,表明该引物的特异性良好。鹅组织PCR产物的核苷酸序列与GenBank中检索到的相应序列基本符合,相似性为98%;PCR检测方法的检测灵敏度为0.01%。[结论]该PCR检测方法特异性强、准确性好、灵敏度高、可重复性好,为明确食品与饲料的成分和来源,预防禽流感的传播提供了有效的分子生物学检测方法。

[ Objective] The purpose,se was to establish a PCR method for detecting goose-derived components. [ Method] With DNAs from 15 animals as templates,PCR reactions were carried out by using specific primers of goose. The PCR products were detected by 1.4% agarose gel electrophoresis and sequenced to detect the specificity and sensitivity of primers. [ Result] The target fragment with leng＇th as 304 bp was amplified from goose samples through PC R reaction. There was nn target fragment in other species such as turkeys,ducks,quail,ostriches, pigeons,chickens, horses,cattle, sheep,pigs,dogs,donkeys,cats, fish as well as the blank control,indicating that the primers had good specificity. The nucleotide sequences of PCR product from goose tissues were identical with the corresponding sequence retrieved in the Genebank and the similarity was 98% ; the detection sensitivity of PCR method was 0.01%. [ Conclusion ] The PCR detection method had characteristics of strong specificity,good accu- racy, high sensitivity and good repeatability, which provided an effective molecular biological detection technique for definiting the composition and source of food and feed and preventing the spread of avian influenza.