title: Development of a real-time quantitative PCR assay using a TaqMan minor groove binder probe for the detection of α-lactalbumin in food.
abstract: Here, we report the development of a real-time PCR assay using a TaqMan minor groove binder (MGB, Genecore, NCBI: AF249896.1, 806-820) probe and primer sets designed to recognize the α-lactalbumin gene from the cow (Bos taurus). We evaluated the efficacy of this assay for detecting and quantifying cow α-lactalbumin in commercial foods. Our results demonstrated that the developed method was highly sensitive and showed high specificity for cow milk, with consistent detection of 0.05 ng of bovine DNA. We tested 42 commercial food samples with or without cow milk listed as an ingredient by using the developed assay. Among the 42 samples, 26 products that listed milk as an ingredient and 3 products might contain milk showed positive signals, whereas the other 9 products that did not contain milk and 4 products that might contain milk tested negative. Therefore, this method could be widely used for the detection of cow milk allergens in food.
authors.affiliation: School of Life Science, Shanghai University, Shanghai 200444, China.
authors.forename: Guan
authors.initials: G
authors.lastname: Xiao
authors.affiliation: School of Life Science, Shanghai University, Shanghai 200444, China.
authors.forename: Cai
authors.initials: C
authors.lastname: Qin
authors.affiliation: School of Life Science, Shanghai University, Shanghai 200444, China.
authors.forename: Zhang
authors.initials: Z
authors.lastname: Wenju
authors.affiliation: School of Life Science, Shanghai University, Shanghai 200444, China. Electronic address: chenqincc@staff.shu.edu.cn.
authors.forename: Chen
authors.initials: C
authors.lastname: Qin
mesh_terms: D000818:Animals; D010367:Arachis; D002417:Cattle; D031211:Corylus; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005502:Food; D005504:Food Analysis; D006041:Goats; D007768:Lactalbumin / Q000032:analysis / Q000235:genetics*; D008892:Milk / Q000737:chemistry*; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary*; D012474:Salmon; D012680:Sensitivity and Specificity; D013025:Glycine max
chemical_list: D017931:DNA Primers; D007768:Lactalbumin
keywords: TaqMan minor groove binder; milk allergen; real-time PCR; α-lactalbumin