title: Development and Application of a Multiplex Real-Time PCR Assay as an Indicator of Potential Allergenicity in Citrus Fruits.
abstract: The effects of tissue type, harvest maturity, and genetic factors on the expression of genes that related to citrus fruit allergies remain poorly understood. In the present study, a multiplex real-time PCR assay was developed to monitor the expression of citrus allergen genes individually with the advantages of much fewer sample requirements and simultaneously multiple target genes detection. Gene specific primer pairs and Taqman probes of three citrus allergen genes Cit s 1.01, Cit s 2.01, and Cit s 3.01 and the house-keeping gene β-actin were designed based on gene sequence differences. The PCR results showed that differential expression patterns were found during the ripening process. The expression levels of Cit s 3.01 were much higher than those of Cit s 1.01 and Cit s 2.01 in both peel and pulp tissues among 10 citrus cultivars. Data suggested that Kao Phuang Pummelo could be safely consumed with a potential low risk in allergenicity. Considering that assessing allergenicity is one of the tests in food safety, this assay might also facilitate the breeding and production of "allergy-friendly" citrus fruits.
authors: Key Laboratory of Horticultural Plant Biology (Ministry of Education), Huazhong Agricultural University , Wuhan 430070, China.
authors: Jinlong
authors: J
authors: Wu
authors: Key Laboratory of Horticultural Plant Biology (Ministry of Education), Huazhong Agricultural University , Wuhan 430070, China.
authors: Lin
authors: L
authors: Chen
authors: Department of Nutritional Sciences, Oklahoma State University , 419 Human Sciences, Stillwater, Oklahoma 74078, United States.
authors: Dingbo
authors: D
authors: Lin
authors: Key Laboratory of Horticultural Plant Biology (Ministry of Education), Huazhong Agricultural University , Wuhan 430070, China.
authors: Zhaocheng
authors: Z
authors: Ma
authors: Key Laboratory of Horticultural Plant Biology (Ministry of Education), Huazhong Agricultural University , Wuhan 430070, China.
authors: Xiuxin
authors: X
authors: Deng
mesh_terms: D000485:Allergens / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D002957:Citrus / Q000235:genetics / Q000276:immunology*; D003257:Consumer Product Safety; D005512:Food Hypersensitivity / Q000276:immunology; D005638:Fruit / Q000235:genetics / Q000276:immunology*; D017343:Genes, Plant; D060885:Multiplex Polymerase Chain Reaction; D010940:Plant Proteins / Q000235:genetics / Q000276:immunology*; D018749:RNA, Plant / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000485:Allergens; D010940:Plant Proteins; D018749:RNA, Plant
keywords: citrus allergen; genetic factors; harvest maturity; multiplex real-time PCR; tissues